Isolated human kinase proteins, nucleic acid molecules encoding human kinase proteins and uses thereof

ABSTRACT

The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the kinase peptides of the present invention. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the kinase peptides, and methods of identifying modulators of the kinase peptides.

FIELD OF THE INVENTION

[0001] The present invention is in the field of kinase proteins that arerelated to the myosin light chain kinase subfamily, recombinant DNAmolecules, and protein production. The present invention specificallyprovides novel peptides and proteins that effect protein phosphorylationand nucleic acid molecules encoding such peptide and protein molecules,all of which are useful in the development of human therapeutics anddiagnostic compositions and methods.

BACKGROUND OF THE INVENTION

[0002] Protein Kinases

[0003] Kinases regulate many different cell proliferation,differentiation, and signaling processes by adding phosphate groups toproteins. Uncontrolled signaling has been implicated in a variety ofdisease conditions including inflammation, cancer, arteriosclerosis, andpsoriasis. Reversible protein phosphorylation is the main strategy forcontrolling activities of eukaryotic cells. It is estimated that morethan 1000 of the 10,000 proteins active in a typical mammalian cell arephosphorylated. The high energy phosphate, which drives activation, isgenerally transferred from adenosine triphosphate molecules (ATP) to aparticular protein by protein kinases and removed from that protein byprotein phosphatases. Phosphorylation occurs in response toextracellular signals (hormones, neurotransmitters, growth anddifferentiation factors, etc), cell cycle checkpoints, and environmentalor nutritional stresses and is roughly analogous to turning on amolecular switch. When the switch goes on, the appropriate proteinkinase activates a metabolic enzyme, regulatory protein, receptor,cytoskeletal protein, ion channel or pump, or transcription factor.

[0004] The kinases comprise the largest known protein group, asuperfamily of enzymes with widely varied functions and specificities.They are usually named after their substrate, their regulatorymolecules, or some aspect of a mutant phenotype. With regard tosubstrates, the protein kinases may be roughly divided into two groups;those that phosphorylate tyrosine residues (protein tyrosine kinases,PTK) and those that phosphorylate serine or threonine residues(serine/threonine kinases, STK). A few protein kinases have dualspecificity and phosphorylate threonine and tyrosine residues. Almostall kinases contain a similar 250-300 amino acid catalytic domain. TheN-terminal domain, which contains subdomains I-IV, generally folds intoa two-lobed structure, which binds and orients the ATP (or GTP) donormolecule. The larger C terminal lobe, which contains subdomains VI A-XI,binds the protein substrate and carries out the transfer of the gammaphosphate from ATP to the hydroxyl group of a serine, threonine, ortyrosine residue. Subdomain V spans the two lobes.

[0005] The kinases may be categorized into families by the differentamino acid sequences (generally between 5 and 100 residues) located oneither side of, or inserted into loops of, the kinase domain. Theseadded amino acid sequences allow the regulation of each kinase as itrecognizes and interacts with its target protein. The primary structureof the kinase domains is conserved and can be further subdivided into 11subdomains. Each of the 11 subdomains contains specific residues andmotifs or patterns of amino acids that are characteristic of thatsubdomain and are highly conserved (Hardie, G. and Hanks, S. (1995) TheProtein Kinase Facts Books, Vol I:7-20 Academic Press, San Diego,Calif.).

[0006] The second messenger dependent protein kinases primarily mediatethe effects of second messengers such as cyclic AMP (cAMP), cyclic GMP,inositol triphosphate, phosphatidylinositol, 3,4,5-triphosphate,cyclic-ADPribose, arachidonic acid, diacylglycerol andcalcium-calmodulin. The cyclic-AMP dependent protein kinases (PKA) areimportant members of the STK family. Cyclic-AMP is an intracellularmediator of hormone action in all prokaryotic and animal cells that havebeen studied. Such hormone-induced cellular responses include thyroidhormone secretion, cortisol secretion, progesterone secretion, glycogenbreakdown, bone resorption, and regulation of heart rate and force ofheart muscle contraction. PKA is found in all animal cells and isthought to account for the effects of cyclic-AMP in most of these cells.Altered PKA expression is implicated in a variety of disorders anddiseases including cancer, thyroid disorders, diabetes, atherosclerosis,and cardiovascular disease (Isselbacher, K. J. et al. (1994) Harrison'sPrinciples of Internal Medicine, McGraw-Hill, New York, N.Y., pp.416-431, 1887).

[0007] Calcium-calmodulin (CaM) dependent protein kinases are alsomembers of STK family. Calmodulin is a calcium receptor that mediatesmany calcium regulated processes by binding to target proteins inresponse to the binding of calcium. The principle target protein inthese processes is CaM dependent protein kinases. CaM-kinases areinvolved in regulation of smooth muscle contraction (MLC kinase),glycogen breakdown (phosphorylase kinase), and neurotransmission (CaMkinase I and CaM kinase II). CaM kinase I phosphorylates a variety ofsubstrates including the neurotransmitter related proteins synapsin Iand II, the gene transcription regulator, CREB, and the cystic fibrosisconductance regulator protein, CFTR (Haribabu, B. et al. (1995) EMBOJournal 14:3679-86). CaM II kinase also phosphorylates synapsin atdifferent sites, and controls the synthesis of catecholamines in thebrain through phosphorylation and activation of tyrosine hydroxylase.Many of the CaM kinases are activated by phosphorylation in addition tobinding to CaM. The kinase may autophosphorylate itself, or bephosphorylated by another kinase as part of a “kinase cascade”.

[0008] Another ligand-activated protein kinase is 5′-AMP-activatedprotein kinase (AMPK) (Gao, G. et al. (1996) J. Biol Chem. 15:8675-81).Mammalian AMPK is a regulator of fatty acid and sterol synthesis throughphosphorylation of the enzymes acetyl-CoA carboxylase andhydroxymethylglutaryl-CoA reductase and mediates responses of thesepathways to cellular stresses such as heat shock and depletion ofglucose and ATP. AMPK is a heterotrimeric complex comprised of acatalytic alpha subunit and two non-catalytic beta and gamma subunitsthat are believed to regulate the activity of the alpha subunit.Subunits of AMPK have a much wider distribution in non-lipogenic tissuessuch as brain, heart, spleen, and lung than expected. This distributionsuggests that its role may extend beyond regulation of lipid metabolismalone.

[0009] The mitogen-activated protein kinases (MAP) are also members ofthe STK family. MAP kinases also regulate intracellular signalingpathways. They mediate signal transduction from the cell surface to thenucleus via phosphorylation cascades. Several subgroups have beenidentified, and each manifests different substrate specificities andresponds to distinct extracellular stimuli (Egan, S. E. and Weinberg, R.A. (1993) Nature 365:781 -783). MAP kinase signaling pathways arepresent in mammalian cells as well as in yeast. The extracellularstimuli that activate mammalian pathways include epidermal growth factor(EGF), ultraviolet light, hyperosmolar medium, heat shock, endotoxiclipopolysaccharide (LPS), and pro-inflammatory cytokines such as tumornecrosis factor (TNF) and interleukin-1 (IL-1).

[0010] PRK (proliferation-related kinase) is a serum/cytokine inducibleSTK that is involved in regulation of the cell cycle and cellproliferation in human megakaroytic cells (Li, B. et al. (1996) J. Biol.Chem. 271:19402-8). PRK is related to the polo (derived from humans pologene) family of STKs implicated in cell division. PRK is downregulatedin lung tumor tissue and may be a proto-oncogene whose deregulatedexpression in normal tissue leads to oncogenic transformation. AlteredMAP kinase expression is implicated in a variety of disease conditionsincluding cancer, inflammation, immune disorders, and disordersaffecting growth and development.

[0011] The cyclin-dependent protein kinases (CDKs) are another group ofSTKs that control the progression of cells through the cell cycle.Cyclins are small regulatory proteins that act by binding to andactivating CDKs that then trigger various phases of the cell cycle byphosphorylating and activating selected proteins involved in the mitoticprocess. CDKs are unique in that they require multiple inputs to becomeactivated. In addition to the binding of cyclin, CDK activation requiresthe phosphorylation of a specific threonine residue and thedephosphorylation of a specific tyrosine residue.

[0012] Protein tyrosine kinases, PTKs, specifically phosphorylatetyrosine residues on their target proteins and may be divided intotransmembrane, receptor PTKs and nontransmembrane, non-receptor PTKs.Transmembrane protein-tyrosine kinases are receptors for most growthfactors. Binding of growth factor to the receptor activates the transferof a phosphate group from ATP to selected tyrosine side chains of thereceptor and other specific proteins. Growth factors (GF) associatedwith receptor PTKs include; epidermal GF, platelet-derived GF,fibroblast GF, hepatocyte GF, insulin and insulin-like GFs, nerve GF,vascular endothelial GF, and macrophage colony stimulating factor.

[0013] Non-receptor PTKs lack transmembrane regions and, instead, formcomplexes with the intracellular regions of cell surface receptors. Suchreceptors that function through non-receptor PTKs include those forcytokines, hormones (growth hormone and prolactin) and antigen-specificreceptors on T and B lymphocytes.

[0014] Many of these PTKs were first identified as the products ofmutant oncogenes in cancer cells where their activation was no longersubject to normal cellular controls. in fact, about one third of theknown oncogenes encode PTKs, and it is well known that cellulartransformation (oncogenesis) is often accompanied by increased tyrosinephosphorylation activity (Carbonneau H and Tonks N K (1992) Annu. Rev.Cell. Biol. 8:463-93). Regulation of PTK activity may therefore be animportant strategy in controlling some types of cancer.

[0015] Myosin Light Chain Kinase

[0016] Activation of smooth/nonmuscle myosin light chain kinase (MLCK)by Ca/calmodulin results in phosphorylation of myosin regulatory lightchain that plays important roles in initiation of smooth musclecontraction, endothelial cell retraction, secretion, and other cellularprocesses (Stull et al., in International Symposium on Regulation of theContractile Cycle in Smooth Muscle, Apr. 26, 1995, Mie, Japan). The samemyosin light chain kinases are present in smooth and nonmuscletissues.(Gallagher et al., J Biol Chem 1991 Dec 15;266(35):23936-44,Published erratum appears in J Biol Chem May 5, 1992;267(13):9450). Thephosphorylation of myosin light chains by myosin light chain kinase is akey event in agonist-mediated endothelial cell gap formation andvascular permeability. Amino acid sequence analysis indicatesendothelial MLCK consensus sequences for a variety of protein kinasesincluding highly conserved potential phosphorylation sites forcAMP-dependent protein kinase A (PKA) in the CaM-binding region.Augmentation of intracellular cAMP levels markedly enhanced MLCKphosphorylation (2.5-fold increase) and reduced kinase activity in MLCKimmunoprecipitates (4-fold decreases) (Garcia et al., Am J Respir CellMol Biol 1997 May;16(5):489-94). The smooth/nonmuscle myosin light chainkinase contains a catalytic core homologous to that of other proteinkinases and a carboxyl-terminal regulatory domain consisting of both aninhibitory sequence and a calmodulin-binding sequence (Kemp et al.,Trends Biochem. Sci. 19, 440-444, 1994; Stull et al., 1995). Initially,inspection of the linear sequence within the regulatory domain revealeda similar number and sequential arrangement of 4 basic residues withthose shown to be important substrate determinants in a syntheticpeptide containing residues 11-23 of the myosin regulatory light chain.Thus, it has been proposed that the regulatory domain contained apseudosubstrate inhibitory sequence whereby 4 specific basic residues inmyosin light chain kinase mimic the basic substrate determinants in thelight chain peptide substrate. Binding of the pseudosubstrate sequenceto the active site inhibited activity. Intrasteric inhibition involvesan autoinhibitory sequence that folds back on the catalytic site toinhibit kinase activity as opposed to an allosteric mechanism whereby aconformational change induced at a site distinct from the active sitewould be responsible for regulation of enzyme activity (Kemp et al.,Biochim. Biophys. Acta. 1094, 67-76, 1991). The sequence comprising thepseudosubstrate region was later expanded to include overlap with thecomplete amino terminus of the light chain (Faux et al., Mol. Cell.Biochem. 128, 81-91, 1993). However, these additional residues (l, 2, 3,4, 5, 6, 7, 8, 9, 10) are not important for substrate binding and thusare not part of the consensus phosphorylation sequence (Kemp et al.,Trends Biochem. Sci. 15, 342-346, 1990).

[0017] Kinase proteins, particularly members of the myosin light chainkinase subfamily, are a major target for drug action and development.Accordingly, it is valuable to the field of pharmaceutical developmentto identify and characterize previously unknown members of thissubfamily of kinase proteins. The present invention advances the stateof the art by providing previously unidentified human kinase proteinsthat have homology to members of the myosin light chain kinasesubfamily.

SUMMARY OF THE INVENTION

[0018] The present invention is based in part on the identification ofamino acid sequences of human kinase peptides and proteins that arerelated to the myosin light chain kinase subfamily, as well as allelicvariants and other mammalian orthologs thereof. These unique peptidesequences, and nucleic acid sequences that encode these peptides, can beused as models for the development of human therapeutic targets, aid inthe identification of therapeutic proteins, and serve as targets for thedevelopment of human therapeutic agents that modulate kinase activity incells and tissues that express the kinase. Experimental data as providedin FIG. 1 indicates expression in the human placenta, kidney, lung,skeletal muscle, heart, fetal brain, and colon carcinoma.

DESCRIPTION OF THE FIGURE SHEETS

[0019]FIG. 1 provides the nucleotide sequence of a cDNA molecule ortranscript sequence that encodes the kinase protein of the presentinvention. (SEQ ID NO: 1) In addition, structure and functionalinformation is provided, such as ATG start, stop and tissuedistribution, where available, that allows one to readily determinespecific uses of inventions based on this molecular sequence.Experimental data as provided in FIG. 1 indicates expression in thehuman placenta, kidney, lung, skeletal muscle, heart, fetal brain, andcolon carcinoma.

[0020]FIG. 2 provides the predicted amino acid sequence of the kinase ofthe present invention. (SEQ ID NO:2) In addition structure andfunctional information such as protein family, function, andmodification sites is provided where available, allowing one to readilydetermine specific uses of inventions based on this molecular sequence.

[0021]FIG. 3 provides genomic sequences that span the gene encoding thekinase protein of the present invention. (SEQ ID NO:3) In additionstructure and functional information, such as intron/exon structure,promoter location, etc., is provided where available, allowing one toreadily determine specific uses of inventions based on this molecularsequence. 6 SNPs, have been identified in the gene encoding the kinaseprotein provided by the present invention and are given in FIG. 3.

DETAILED DESCRIPTION OF THE INVENTION

[0022] General Description

[0023] The present invention is based on the sequencing of the humangenome. During the sequencing and assembly of the human genome, analysisof the sequence information revealed previously unidentified fragmentsof the human genome that encode peptides that share structural and/orsequence homology to protein/peptide/domains identified andcharacterized within the art as being a kinase protein or part of akinase protein and are related to the myosin light chain kinasesubfamily. Utilizing these sequences, additional genomic sequences wereassembled and transcript and/or cDNA sequences were isolated andcharacterized. Based on this analysis, the present invention providesamino acid sequences of human kinase peptides and proteins that arerelated to the myosin light chain kinase subfamily, nucleic acidsequences in the form of transcript sequences, cDNA sequences and/orgenomic sequences that encode these kinase peptides and proteins,nucleic acid variation (allelic information), tissue distribution ofexpression, and information about the closest art knownprotein/peptide/domain that has structural or sequence homology to thekinase of the present invention.

[0024] In addition to being previously unknown, the peptides that areprovided in the present invention are selected based on their ability tobe used for the development of commercially important products andservices. Specifically, the present peptides are selected based onhomology and/or structural relatedness to known kinase proteins of themyosin light chain kinase subfamily and the expression pattern observed.Experimental data as provided in FIG. 1 indicates expression in thehuman placenta, kidney, lung, skeletal muscle, heart, fetal brain, andcolon carcinoma. The art has clearly established the commercialimportance of members of this family of proteins and proteins that haveexpression patterns similar to that of the present gene. Some of themore specific features of the peptides of the present invention, and theuses thereof, are described herein, particularly in the Background ofthe Invention and in the annotation provided in the Figures, and/or areknown within the art for each of the known myosin light chain kinasefamily or subfamily of kinase proteins.

[0025] Specific Embodiments

[0026] Peptide Molecules

[0027] The present invention provides nucleic acid sequences that encodeprotein molecules that have been identified as being members of thekinase family of proteins and are related to the myosin light chainkinase subfamily (protein sequences are provided in FIG. 2,transcript/cDNA sequences are provided in FIG. 1 and genomic sequencesare provided in FIG. 3). The peptide sequences provided in FIG. 2, aswell as the obvious variants described herein, particularly allelicvariants as identified herein and using the information in FIG. 3, willbe referred herein as the kinase peptides of the present invention,kinase peptides, or peptides/proteins of the present invention.

[0028] The present invention provides isolated peptide and proteinmolecules that consist of, consist essentially of, or comprise the aminoacid sequences of the kinase peptides disclosed in the FIG. 2, (encodedby the nucleic acid molecule shown in FIG. 1, transcript/cDNA or FIG. 3,genomic sequence), as well as all obvious variants of these peptidesthat are within the art to make and use. Some of these variants aredescribed in detail below.

[0029] As used herein, a peptide is said to be “isolated” or “purified”when it is substantially free of cellular material or free of chemicalprecursors or other chemicals. The peptides of the present invention canbe purified to homogeneity or other degrees of purity. The level ofpurification will be based on the intended use. The critical feature isthat the preparation allows for the desired function of the peptide,even if in the presence of considerable amounts of other components (thefeatures of an isolated nucleic acid molecule is discussed below).

[0030] In some uses, “substantially free of cellular material” includespreparations of the peptide having less than about 30% (by dry weight)other proteins (i.e., contaminating protein), less than about 20% otherproteins, less than about 10% other proteins, or less than about 5%other proteins. When the peptide is recombinantly produced, it can alsobe substantially free of culture medium, i.e., culture medium representsless than about 20% of the volume of the protein preparation.

[0031] The language “substantially free of chemical precursors or otherchemicals” includes preparations of the peptide in which it is separatedfrom chemical precursors or other chemicals that are involved in itssynthesis. In one embodiment, the language “substantially free ofchemical precursors or other chemicals” includes preparations of thekinase peptide having less than about 30% (by dry weight) chemicalprecursors or other chemicals, less than about 20% chemical precursorsor other chemicals, less than about 10% chemical precursors or otherchemicals, or less than about 5% chemical precursors or other chemicals.

[0032] The isolated kinase peptide can be purified from cells thatnaturally express it, purified from cells that have been altered toexpress it (recombinant), or synthesized using known protein synthesismethods. Experimental data as provided in FIG. 1 indicates expression inthe human placenta, kidney, lung, skeletal muscle, heart, fetal brain,and colon carcinoma. For example, a nucleic acid molecule encoding thekinase peptide is cloned into an expression vector, the expressionvector introduced into a host cell and the protein expressed in the hostcell. The protein can then be isolated from the cells by an appropriatepurification scheme using standard protein purification techniques. Manyof these techniques are described in detail below.

[0033] Accordingly, the present invention provides proteins that consistof the amino acid sequences provided in FIG. 2 (SEQ NO:2), for example,proteins encoded by the transcript/cDNA nucleic acid sequences shown inFIG. 1 (SEQ ID NO: 1) and the genomic sequences provided in FIG. 3 (SEQID NO:3). The amino acid sequence of such a protein is provided in FIG.2. A protein consists of an amino acid sequence when the amino acidsequence is the final amino acid sequence of the protein.

[0034] The present invention further provides proteins that consistessentially of the amino acid sequences provided in FIG. 2 (SEQ IDNO:2), for example, proteins encoded by the transcript/cDNA nucleic acidsequences shown in FIG. 1 (SEQ ID NO: 1) and the genomic sequencesprovided in FIG. 3 (SEQ ID NO:3). A protein consists essentially of anamino acid sequence when such an amino acid sequence is present withonly a few additional amino acid residues, for example from about 1 toabout 100 or so additional residues, typically from 1 to about 20additional residues in the final protein.

[0035] The present invention further provides proteins that comprise theamino acid sequences provided in FIG. 2 (SEQ ID NO:2), for example,proteins encoded by the transcript/cDNA nucleic acid sequences shown inFIG. 1 (SEQ ID NO:1) and the genomic sequences provided in FIG. 3 (SEQID NO:3). A protein comprises an amino acid sequence when the amino acidsequence is at least part of the final amino acid sequence of theprotein. In such a fashion, the protein can be only the peptide or haveadditional amino acid molecules, such as amino acid residues (contiguousencoded sequence) that are naturally associated with it or heterologousamino acid residues/peptide sequences. Such a protein can have a fewadditional amino acid residues or can comprise several hundred or moreadditional amino acids. The preferred classes of proteins that arecomprised of the kinase peptides of the present invention are thenaturally occurring mature proteins. A brief description of how varioustypes of these proteins can be made/isolated is provided below.

[0036] The kinase peptides of the present invention can be attached toheterologous sequences to form chimeric or fusion proteins. Suchchimeric and fusion proteins comprise a kinase peptide operativelylinked to a heterologous protein having an amino acid sequence notsubstantially homologous to the kinase peptide. “Operatively linked”indicates that the kinase peptide and the heterologous protein are fusedin-frame. The heterologous protein can be fused to the N-terminus orC-terminus of the kinase peptide.

[0037] In some uses, the fusion protein does not affect the activity ofthe kinase peptide per se. For example, the fusion protein can include,but is not limited to, enzymatic fusion proteins, for examplebeta-galactosidase fusions, yeast two-hybrid GAL fusions, poly-Hisfusions, MYC-tagged, HI-tagged and Ig fusions. Such fusion proteins,particularly poly-His fusions, can facilitate the purification ofrecombinant kinase peptide. In certain host cells (e.g., mammalian hostcells), expression and/or secretion of a protein can be increased byusing a heterologous signal sequence.

[0038] A chimeric or fusion protein can be produced by standardrecombinant DNA techniques. For example, DNA fragments coding for thedifferent protein sequences are ligated together in-frame in accordancewith conventional techniques. In another embodiment, the fusion gene canbe synthesized by conventional techniques including automated DNAsynthesizers. Alternatively, PCR amplification of gene fragments can becarried out using anchor primers which give rise to complementaryoverhangs between two consecutive gene fragments which can subsequentlybe annealed and re-amplified to generate a chimeric gene sequence (seeAusubel et al., Current Protocols in Molecular Biology, 1992). Moreover,many expression vectors are commercially available that already encode afusion moiety (e.g., a GST protein). A kinase peptide-encoding nucleicacid can be cloned into such an expression vector such that the fusionmoiety is linked in-frame to the kinase peptide.

[0039] As mentioned above, the present invention also provides andenables obvious variants of the amino acid sequence of the proteins ofthe present invention, such as naturally occurring mature forms of thepeptide, allelic/sequence variants of the peptides, non-naturallyoccurring recombinantly derived variants of the peptides, and orthologsand paralogs of the peptides. Such variants can readily be generatedusing art-known techniques in the fields of recombinant nucleic acidtechnology and protein biochemistry. It is understood, however, thatvariants exclude any amino acid sequences disclosed prior to theinvention.

[0040] Such variants can readily be identified/made using moleculartechniques and the sequence information disclosed herein. Further, suchvariants can readily be distinguished from other peptides based onsequence and/or structural homology to the kinase peptides of thepresent invention. The degree of homology/identity present will be basedprimarily on whether the peptide is a functional variant ornon-functional variant, the amount of divergence present in the paralogfamily and the evolutionary distance between the orthologs.

[0041] To determine the percent identity of two amino acid sequences ortwo nucleic acid sequences, the sequences are aligned for optimalcomparison purposes (e.g., gaps can be introduced in one or both of afirst and a second amino acid or nucleic acid sequence for optimalalignment and non-homologous sequences can be disregarded for comparisonpurposes). In a preferred embodiment, at least 30%, 40%, 50%, 60%, 70%,80%, or 90% or more of the length of a reference sequence is aligned forcomparison purposes. The amino acid residues or nucleotides atcorresponding amino acid positions or nucleotide positions are thencompared. When a position in the first sequence is occupied by the sameamino acid residue or nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position (asused herein amino acid or nucleic acid “identity” is equivalent to aminoacid or nucleic acid “homology”). The percent identity between the twosequences is a function of the number of identical positions shared bythe sequences, taking into account the number of gaps, and the length ofeach gap, which need to be introduced for optimal alignment of the twosequences.

[0042] The comparison of sequences and determination of percent identityand similarity between two sequences can be accomplished using amathematical algorithm. (Computational Molecular Biology, Lesk, A. M.,ed., Oxford University Press, New York, 1988; Biocomputing: Informaticsand Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993;Computer Analysis of Sequence Data, Part 1, Griffm, A. M., and Griffin,H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis inMolecular Biology, von Heinje, G., Academic Press, 1987; and SequenceAnalysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press,New York, 1991). In a preferred embodiment, the percent identity betweentwo amino acid sequences is determined using the Needleman and Wunsch(J. Mol. Biol. (48):444-453 (1970)) algorithm which has beenincorporated into the GAP program in the GCG software package (availableat http://www.gcg.com), using either a Blossom 62 matrix or a PAM250matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a lengthweight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, thepercent identity between two nucleotide sequences is determined usingthe GAP program in the GCG software package (Devereux, J., et al.,Nucleic Acids Res. 12(1):387 (1984)) (available at http://www.gcg.com),using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80and a length weight of 1, 2, 3, 4, 5, or 6. In another embodiment, thepercent identity between two amino acid or nucleotide sequences isdetermined using the algorithm of E. Myers and W. Miller (CABIOS,4:11-17 (1989)) which has been incorporated into the ALIGN program(version 2.0), using a PAM120 weight residue table, a gap length penaltyof 12 and a gap penalty of 4.

[0043] The nucleic acid and protein sequences of the present inventioncan further be used as a “query sequence” to perform a search againstsequence databases to, for example, identify other family members orrelated sequences. Such searches can be performed using the NBLAST andXBLAST programs (version 2.0) of Altschul, et al. (J. Mol. Biol.215:403-10 (1990)). BLAST nucleotide searches can be performed with theNBLAST program, score=100, wordlength=12 to obtain nucleotide sequenceshomologous to the nucleic acid molecules of the invention. BLAST proteinsearches can be performed with the XBLAST program, score=50,wordlength=3 to obtain amino acid sequences homologous to the proteinsof the invention. To obtain gapped alignments for comparison purposes,Gapped BLAST can be utilized as described in Altschul et al. (NucleicAcids Res. 25(17):3389-3402 (1997)). When utilizing BLAST and gappedBLAST programs, the default parameters of the respective programs (e.g.,XBLAST and NBLAST) can be used.

[0044] Full-length pre-processed forms, as well as mature processedforms, of proteins that comprise one of the peptides of the presentinvention can readily be identified as having complete sequence identityto one of the kinase peptides of the present invention as well as beingencoded by the same genetic locus as the kinase peptide provided herein.As indicated by the data presented in FIG. 3, the map position wasdetermined to be on chromosome 1 by ePCR, and confirmed with radiationhybrid mapping. As indicated by the data presented in FIG. 3, the geneprovided by the present invention encoding a novel phosphatase maps topublic BAC AC AC023889, which is known to be located on human chromosome1.

[0045] Allelic variants of a kinase peptide can readily be identified asbeing a human protein having a high degree (significant) of sequencehomology/identity to at least a portion of the kinase peptide as well asbeing encoded by the same genetic locus as the kinase peptide providedherein. Genetic locus can readily be determined based on the genomicinformation provided in FIG. 3, such as the genomic sequence mapped tothe reference human. As indicated by the data presented in FIG. 3, themap position was determined to be on chromosome 1 by ePCR, and confirmedwith radiation hybrid mapping. As indicated by the data presented inFIG. 3, the gene provided by the present invention encoding a novelphosphatase maps to public BAC AC AC023889, which is known to be locatedon human chromosome 1. As used herein, two proteins (or a region of theproteins) have significant homology when the amino acid sequences aretypically at least about 70-80%, 80-90%, and more typically at leastabout 90-95% or more homologous. A significantly homologous amino acidsequence, according to the present invention, will be encoded by anucleic acid sequence that will hybridize to a kinase peptide encodingnucleic acid molecule under stringent conditions as more fully describedbelow.

[0046]FIG. 3 provides information on SNPs that have been identified in agene encoding the kinase protein of the present invention. 6 SNPvariants were found, and all SNPs in exons, of which 3 of these causechanges in the amino acid sequence (i.e., nonsynonymous SNPs). Thechanges in the amino acid sequence that these SNPs cause is indicated inFIG. 3 and can readily be determined using the universal genetic codeand the protein sequence provided in FIG. 2 as a reference.

[0047] Paralogs of a kinase peptide can readily be identified as havingsome degree of significant sequence homology/identity to at least aportion of the kinase peptide, as being encoded by a gene from humans,and as having similar activity or function. Two proteins will typicallybe considered paralogs when the amino acid sequences are typically atleast about 60% or greater, and more typically at least about 70% orgreater homology through a given region or domain. Such paralogs will beencoded by a nucleic acid sequence that will hybridize to a kinasepeptide encoding nucleic acid molecule under moderate to stringentconditions as more fully described below.

[0048] Orthologs of a kinase peptide can readily be identified as havingsome degree of significant sequence homology/identity to at least aportion of the kinase peptide as well as being encoded by a gene fromanother organism. Preferred orthologs will be isolated from mammals,preferably primates, for the development of human therapeutic targetsand agents. Such orthologs will be encoded by a nucleic acid sequencethat will hybridize to a kinase peptide encoding nucleic acid moleculeunder moderate to stringent conditions, as more fully described below,depending on the degree of relatedness of the two organisms yielding theproteins.

[0049] Non-naturally occurring variants of the kinase peptides of thepresent invention can readily be generated using recombinant techniques.Such variants include, but are not limited to deletions, additions andsubstitutions in the amino acid sequence of the kinase peptide. Forexample, one class of substitutions are conserved amino acidsubstitution. Such substitutions are those that substitute a given aminoacid in a kinase peptide by another amino acid of like characteristics.Typically seen as conservative substitutions are the replacements, onefor another, among the aliphatic amino acids Ala, Val, Leu, and Ile;interchange of the hydroxyl residues Ser and Thr; exchange of the acidicresidues Asp and Glu; substitution between the amide residues Asn andGln; exchange of the basic residues Lys and Arg; and replacements amongthe aromatic residues Phe and Tyr. Guidance concerning which amino acidchanges are likely to be phenotypically silent are found in Bowie etal., Science 247:1306-1310 (1990).

[0050] Variant kinase peptides can be fully functional or can lackfunction in one or more activities, e.g. ability to bind substrate,ability to phosphorylate substrate, ability to mediate signaling, etc.Fully functional variants typically contain only conservative variationor variation in non-critical residues or in non-critical regions. FIG. 2provides the result of protein analysis and can be used to identifycritical domains/regions. Functional variants can also containsubstitution of similar amino acids that result in no change or aninsignificant change in function. Alternatively, such substitutions maypositively or negatively affect function to some degree.

[0051] Non-functional variants typically contain one or morenon-conservative amino acid substitutions, deletions, insertions,inversions, or truncation or a substitution, insertion, inversion, ordeletion in a critical residue or critical region.

[0052] Amino acids that are essential for function can be identified bymethods known in the art, such as site-directed mutagenesis oralanine-scanning mutagenesis (Cunningham et al., Science 244:1081-1085(1989)), particularly using the results provided in FIG. 2. The latterprocedure introduces single alanine mutations at every residue in themolecule. The resulting mutant molecules are then tested for biologicalactivity such as kinase activity or in assays such as an in vitroproliferative activity. Sites that are critical for bindingpartner/substrate binding can also be determined by structural analysissuch as crystallization, nuclear magnetic resonance or photoaffinitylabeling (Smith et al., J. Mol. Biol. 224:899-904 (1992); de Vos et al.Science 255:306-312 (1992)).

[0053] The present invention further provides fragments of the kinasepeptides, in addition to proteins and peptides that comprise and consistof such fragments, particularly those comprising the residues identifiedin FIG. 2. The fragments to which the invention pertains, however, arenot to be construed as encompassing fragments that may be disclosedpublicly prior to the present invention.

[0054] As used herein, a fragment comprises at least 8, 10, 12, 14, 16,or more contiguous amino acid residues from a kinase peptide. Suchfragments can be chosen based on the ability to retain one or more ofthe biological activities of the kinase peptide or could be chosen forthe ability to perform a function, e.g. bind a substrate or act as animmunogen. Particularly important fragments are biologically activefragments, peptides that are, for example, about 8 or more amino acidsin length. Such fragments will typically comprise a domain or motif ofthe kinase peptide, e.g., active site, a transmembrane domain or asubstrate-binding domain. Further, possible fragments include, but arenot limited to, domain or motif containing fragments, soluble peptidefragments, and fragments containing immunogenic structures. Predicteddomains and functional sites are readily identifiable by computerprograms well known and readily available to those of skill in the art(e.g., PROSITE analysis). The results of one such analysis are providedin FIG. 2.

[0055] Polypeptides often contain amino acids other than the 20 aminoacids commonly referred to as the 20 naturally occurring amino acids.Further, many amino acids, including the terminal amino acids, may bemodified by natural processes, such as processing and otherpost-translational modifications, or by chemical modification techniqueswell known in the art. Common modifications that occur naturally inkinase peptides are described in basic texts, detailed monographs, andthe research literature, and they are well known to those of skill inthe art (some of these features are identified in FIG. 2).

[0056] Known modifications include, but are not limited to, acetylation,acylation, ADP-ribosylation, amidation, covalent attachment of flavin,covalent attachment of a heme moiety, covalent attachment of anucleotide or nucleotide derivative, covalent attachment of a lipid orlipid derivative, covalent attachment of phosphotidylinositol,cross-linking, cyclization, disulfide bond formation, demethylation,formation of covalent crosslinks, formation of cystine, formation ofpyroglutamate, formylation, gamma carboxylation, glycosylation, GPIanchor formation, hydroxylation, iodination, methylation,myristoylation, oxidation, proteolytic processing, phosphorylation,prenylation, racemization, selenoylation, sulfation, transfer-RNAmediated addition of amino acids to proteins such as arginylation, andubiquitination.

[0057] Such modifications are well known to those of skill in the artand have been described in great detail in the scientific literature.Several particularly common modifications, glycosylation, lipidattachment, sulfation, gamma-carboxylation of glutamic acid residues,hydroxylation and ADP-ribosylation, for instance, are described in mostbasic texts, such as Proteins—Structure and Molecular Properties, 2ndEd., T. E. Creighton, W.H. Freeman and Company, New York (1993). Manydetailed reviews are available on this subject, such as by Wold, F.,Posttranslational Covalent Modification of Proteins, B. C. Johnson, Ed.,Academic Press, New York 1-12 (1983); Seifter et al. (Meth. Enzymol.182: 626-646 (1990)) and Rattan et al. (Ann. N.Y Acad. Sci. 663:48-62(1992)).

[0058] Accordingly, the kinase peptides of the present invention alsoencompass derivatives or analogs in which a substituted amino acidresidue is not one encoded by the genetic code, in which a substituentgroup is included, in which the mature kinase peptide is fused withanother compound, such as a compound to increase the half-life of thekinase peptide (for example, polyethylene glycol), or in which theadditional amino acids are fused to the mature kinase peptide, such as aleader or secretory sequence or a sequence for purification of themature kinase peptide or a pro-protein sequence.

[0059] Protein/Peptide Uses

[0060] The proteins of the present invention can be used in substantialand specific assays related to the functional information provided inthe Figures; to raise antibodies or to elicit another immune response;as a reagent (including the labeled reagent) in assays designed toquantitatively determine levels of the protein (or its binding partneror ligand) in biological fluids; and as markers for tissues in which thecorresponding protein is preferentially expressed (either constitutivelyor at a particular stage of tissue differentiation or development or ina disease state). Where the protein binds or potentially binds toanother protein or ligand (such as, for example, in a kinase-effectorprotein interaction or kinase-ligand interaction), the protein can beused to identify the binding partner/ligand so as to develop a system toidentify inhibitors of the binding interaction. Any or all of these usesare capable of being developed into reagent grade or kit format forcommercialization as commercial products.

[0061] Substantial chemical and structural homology exists between thekinase protein of the present invention described herein and myosinlight chain kinase (see FIG. 1). As discussed in the background, myosinlight chain kinase are known in the art to be involved in smooth musclecontraction, endothelial cell retraction, secretion, and other cellularprocess. Accordingly, the myosin light chain kinase, and the encodinggene, provided by the present invention is useful for treating,preventing, and/or diagnosing disorders associated with muscle,endothelial cells.

[0062] Methods for performing the uses listed above are well known tothose skilled in the art. References disclosing such methods include“Molecular Cloning: A Laboratory Manual”, 2d ed., Cold Spring HarborLaboratory Press, Sambrook, J., E. F. Fritsch and T. Maniatis eds.,1989, and “Methods in Enzymology: Guide to Molecular CloningTechniques”, Academic Press, Berger, S. L. and A. R. Kimmel eds., 1987.

[0063] The potential uses of the peptides of the present invention arebased primarily on the source of the protein as well as the class/actionof the protein. For example, kinases isolated from humans and theirhuman/mammalian orthologs serve as targets for identifying agents foruse in mammalian therapeutic applications, e.g. a human drug,particularly in modulating a biological or pathological response in acell or tissue that expresses the kinase. Experimental data as providedin FIG. 1 indicates that kinase proteins of the present invention areexpressed in the human placenta, kidney, lung, skeletal muscle, heart,fetal brain, and colon carcinoma. Specifically, a virtual northern blotshows expression in human colon carcinoma. In addition, PCR-based tissuescreening panel indicates expression in human placenta, kidney, lung,skeletal muscle, heart, and fetal brain. A large percentage ofpharmaceutical agents are being developed that modulate the activity ofkinase proteins, particularly members of the myosin light chain kinasesubfamily (see Background of the Invention). The structural andfunctional information provided in the Background and Figures providespecific and substantial uses for the molecules of the presentinvention, particularly in combination with the expression informationprovided in FIG. 1. Experimental data as provided in FIG. 1 indicatesexpression in the human placenta, kidney, lung, skeletal muscle, heart,fetal brain, and colon carcinoma. Such uses can readily be determinedusing the information provided herein, that which is known in the art,and routine experimentation.

[0064] The proteins of the present invention (including variants andfragments that may have been disclosed prior to the present invention)are useful for biological assays related to kinases that are related tomembers of the myosin light chain kinase subfamily. Such assays involveany of the known kinase functions or activities or properties useful fordiagnosis and treatment of kinase-related conditions that are specificfor the subfamily of kinases that the one of the present inventionbelongs to, particularly in cells and tissues that express the kinase.Experimental data as provided in FIG. 1 indicates that kinase proteinsof the present invention are expressed in the human placenta, kidney,lung, skeletal muscle, heart, fetal brain, and colon carcinoma.Specifically, a virtual northern blot shows expression in human coloncarcinoma. In addition, PCR-based tissue screening panel indicatesexpression in human placenta, kidney, lung, skeletal muscle, heart, andfetal brain.

[0065] The proteins of the present invention are also useful in drugscreening assays, in cell-based or cell-free systems. Cell-based systemscan be native, i.e., cells that normally express the kinase, as a biopsyor expanded in cell culture. Experimental data as provided in FIG. 1indicates expression in the human placenta, kidney, lung, skeletalmuscle, heart, fetal brain, and colon carcinoma. In an alternateembodiment, cell-based assays involve recombinant host cells expressingthe kinase protein.

[0066] The polypeptides can be used to identify compounds that modulatekinase activity of the protein in its natural state or an altered formthat causes a specific disease or pathology associated with the kinase.Both the kinases of the present invention and appropriate variants andfragments can be used in high-throughput screens to assay candidatecompounds for the ability to bind to the kinase. These compounds can befurther screened against a functional kinase to determine the effect ofthe compound on the kinase activity. Further, these compounds can betested in animal or invertebrate systems to determineactivity/effectiveness. Compounds can be identified that activate(agonist) or inactivate (antagonist) the kinase to a desired degree.

[0067] Further, the proteins of the present invention can be used toscreen a compound for the ability to stimulate or inhibit interactionbetween the kinase protein and a molecule that normally interacts withthe kinase protein, e.g. a substrate or a component of the signalpathway that the kinase protein normally interacts (for example, anotherkinase). Such assays typically include the steps of combining the kinaseprotein with a candidate compound under conditions that allow the kinaseprotein, or fragment, to interact with the target molecule, and todetect the formation of a complex between the protein and the target orto detect the biochemical consequence of the interaction with the kinaseprotein and the target, such as any of the associated effects of signaltransduction such as protein phosphorylation, cAMP turnover, andadenylate cyclase activation, etc.

[0068] Candidate compounds include, for example, 1) peptides such assoluble peptides, including Ig-tailed fusion peptides and members ofrandom peptide libraries (see, e.g., Lam et al., Nature 354:82-84(1991); Houghten et al., Nature 354:84-86 (1991)) and combinatorialchemistry-derived molecular libraries made of D- and/or L-configurationamino acids; 2) phosphopeptides (e.g., members of random and partiallydegenerate, directed phosphopeptide libraries, see, e.g., Songyang etal., Cell 72:767-778 (1993)); 3) antibodies (e.g., polyclonal,monoclonal, humanized, anti-idiotypic, chimeric, and single chainantibodies as well as Fab, F(ab′)₂, Fab expression library fragments,and epitope-binding fragments of antibodies); and 4) small organic andinorganic molecules (e.g., molecules obtained from combinatorial andnatural product libraries).

[0069] One candidate compound is a soluble fragment of the receptor thatcompetes for substrate binding. Other candidate compounds include mutantkinases or appropriate fragments containing mutations that affect kinasefunction and thus compete for substrate. Accordingly, a fragment thatcompetes for substrate, for example with a higher affinity, or afragment that binds substrate but does not allow release, is encompassedby the invention.

[0070] The invention further includes other end point assays to identifycompounds that modulate (stimulate or inhibit) kinase activity. Theassays typically involve an assay of events in the signal transductionpathway that indicate kinase activity. Thus, the phosphorylation of asubstrate, activation of a protein, a change in the expression of genesthat are up- or down-regulated in response to the kinase proteindependent signal cascade can be assayed.

[0071] Any of the biological or biochemical functions mediated by thekinase can be used as an endpoint assay. These include all of thebiochemical or biochemical/biological events described herein, in thereferences cited herein, incorporated by reference for these endpointassay targets, and other functions known to those of ordinary skill inthe art or that can be readily identified using the information providedin the Figures, particularly FIG. 2. Specifically, a biological functionof a cell or tissues that expresses the kinase can be assayed.Experimental data as provided in FIG. 1 indicates that kinase proteinsof the present invention are expressed in the human placenta, kidney,lung, skeletal muscle, heart, fetal brain, and colon carcinoma.Specifically, a virtual northern blot shows expression in human coloncarcinoma. In addition, PCR-based tissue screening panel indicatesexpression in human placenta, kidney, lung, skeletal muscle, heart, andfetal brain.

[0072] Binding and/or activating compounds can also be screened by usingchimeric kinase proteins in which the amino terminal extracellulardomain, or parts thereof, the entire transmembrane domain or subregions,such as any of the seven transmembrane segments or any of theintracellular or extracellular loops and the carboxy terminalintracellular domain, or parts thereof, can be replaced by heterologousdomains or subregions. For example, a substrate-binding region can beused that interacts with a different substrate then that which isrecognized by the native kinase. Accordingly, a different set of signaltransduction components is available as an end-point assay foractivation. This allows for assays to be performed in other than thespecific host cell from which the kinase is derived.

[0073] The proteins of the present invention are also useful incompetition binding assays in methods designed to discover compoundsthat interact with the kinase (e.g. binding partners and/or ligands).Thus, a compound is exposed to a kinase polypeptide under conditionsthat allow the compound to bind or to otherwise interact with thepolypeptide. Soluble kinase polypeptide is also added to the mixture. Ifthe test compound interacts with the soluble kinase polypeptide, itdecreases the amount of complex or mild or activity from the kinasetarget. This type of assay is particularly useful in cases in whichcompounds are sought that interact with specific regions of the kinase.Thus, the soluble polypeptide that competes with the target kinaseregion is designed to contain peptide sequences corresponding to theregion of interest.

[0074] To perform cell free drug screening assays, it is sometimesdesirable to immobilize either the kinase protein, or fragment, or itstarget molecule to facilitate separation of complexes from uncomplexedforms of one or both of the proteins, as well as to accommodateautomation of the assay.

[0075] Techniques for immobilizing proteins on matrices can be used inthe drug screening assays. In one embodiment, a fusion protein can beprovided which adds a domain that allows the protein to be bound to amatrix. For example, glutathione-S-transferase fusion proteins can beadsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis,Mo.) or glutathione derivatized microtitre plates, which are thencombined with the cell lysates (e.g., ³⁵S-labeled) and the candidatecompound, and the mixture incubated under conditions conducive tocomplex formation (e.g., at physiological conditions for salt and pH).Following incubation, the beads are washed to remove any unbound label,and the matrix immobilized and radiolabel determined directly, or in thesupernatant after the complexes are dissociated. Alternatively, thecomplexes can be dissociated from the matrix, separated by SDS-PAGE, andthe level of kinase-binding protein found in the bead fractionquantitated from the gel using standard electrophoretic techniques. Forexample, either the polypeptide or its target molecule can beimmobilized utilizing conjugation of biotin and streptavidin usingtechniques well known in the art. Alternatively, antibodies reactivewith the protein but which do not interfere with binding of the proteinto its target molecule can be derivatized to the wells of the plate, andthe protein trapped in the wells by antibody conjugation. Preparationsof a kinase-binding protein and a candidate compound are incubated inthe kinase protein-presenting wells and the amount of complex trapped inthe well can be quantitated. Methods for detecting such complexes, inaddition to those described above for the GST-immobilized complexes,include immunodetection of complexes using antibodies reactive with thekinase protein target molecule, or which are reactive with kinaseprotein and compete with the target molecule, as well as enzyme-linkedassays which rely on detecting an enzymatic activity associated with thetarget molecule.

[0076] Agents that modulate one of the kinases of the present inventioncan be identified using one or more of the above assays, alone or incombination. It is generally preferable to use a cell-based or cell freesystem first and then confirm activity in an animal or other modelsystem. Such model systems are well known in the art and can readily beemployed in this context.

[0077] Modulators of kinase protein activity identified according tothese drug screening assays can be used to treat a subject with adisorder mediated by the kinase pathway, by treating cells or tissuesthat express the kinase. Experimental data as provided in FIG. 1indicates expression in the human placenta, kidney, lung, skeletalmuscle, heart, fetal brain, and colon carcinoma. These methods oftreatment include the steps of administering a modulator of kinaseactivity in a pharmaceutical composition to a subject in need of suchtreatment, the modulator being identified as described herein.

[0078] In yet another aspect of the invention, the kinase proteins canbe used as “bait proteins” in a two-hybrid assay or three-hybrid assay(see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993) Cell72:223-232; Madura et al. (1993) J. Biol. Chem. 268:12046-12054; Bartelet al. (1993) Biotechniques 14:920-924; Iwabuchi et al. (1993) Oncogene8:1693-1696; and Brent WO94/10300), to identify other proteins, whichbind to or interact with the kinase and are involved in kinase activity.Such kinase-binding proteins are also likely to be involved in thepropagation of signals by the kinase proteins or kinase targets as, forexample, downstream elements of a kinase-mediated signaling pathway.Alternatively, such kinase-binding proteins are likely to be kinaseinhibitors.

[0079] The two-hybrid system is based on the modular nature of mosttranscription factors, which consist of separable DNA-binding andactivation domains. Briefly, the assay utilizes two different DNAconstructs. In one construct, the gene that codes for a kinase proteinis fused to a gene encoding the DNA binding domain of a knowntranscription factor (e.g., GAL-4). In the other construct, a DNAsequence, from a library of DNA sequences, that encodes an unidentifiedprotein (“prey” or “sample”) is fused to a gene that codes for theactivation domain of the known transcription factor. If the “bait” andthe “prey” proteins are able to interact, in vivo, forming akinase-dependent complex, the DNA-binding and activation domains of thetranscription factor are brought into close proximity. This proximityallows transcription of a reporter gene (e.g., LacZ) which is operablylinked to a transcriptional regulatory site responsive to thetranscription factor. Expression of the reporter gene can be detectedand cell colonies containing the functional transcription factor can beisolated and used to obtain the cloned gene which encodes the proteinwhich interacts with the kinase protein.

[0080] This invention further pertains to novel agents identified by theabove-described screening assays. Accordingly, it is within the scope ofthis invention to further use an agent identified as described herein inan appropriate animal model. For example, an agent identified asdescribed herein (e.g., a kinase-modulating agent, an antisense kinasenucleic acid molecule, a kinase-specific antibody, or a kinase-bindingpartner) can be used in an animal or other model to determine theefficacy, toxicity, or side effects of treatment with such an agent.Alternatively, an agent identified as described herein can be used in ananimal or other model to determine the mechanism of action of such anagent. Furthermore, this invention pertains to uses of novel agentsidentified by the above-described screening assays for treatments asdescribed herein.

[0081] The kinase proteins of the present invention are also useful toprovide a target for diagnosing a disease or predisposition to diseasemediated by the peptide. Accordingly, the invention provides methods fordetecting the presence, or levels of, the protein (or encoding mRNA) ina cell, tissue, or organism. Experimental data as provided in FIG. 1indicates expression in the human placenta, kidney, lung, skeletalmuscle, heart, fetal brain, and colon carcinoma. The method involvescontacting a biological sample with a compound capable of interactingwith the kinase protein such that the interaction can be detected. Suchan assay can be provided in a single detection format or amulti-detection format such as an antibody chip array.

[0082] One agent for detecting a protein in a sample is an antibodycapable of selectively binding to protein. A biological sample includestissues, cells and biological fluids isolated from a subject, as well astissues, cells and fluids present within a subject.

[0083] The peptides of the present invention also provide targets fordiagnosing active protein activity, disease, or predisposition todisease, in a patient having a variant peptide, particularly activitiesand conditions that are known for other members of the family ofproteins to which the present one belongs. Thus, the peptide can beisolated from a biological sample and assayed for the presence of agenetic mutation that results in aberrant peptide. This includes aminoacid substitution, deletion, insertion, rearrangement, (as the result ofaberrant splicing events), and inappropriate post-translationalmodification. Analytic methods include altered electrophoretic mobility,altered tryptic peptide digest, altered kinase activity in cell-based orcell-free assay, alteration in substrate or antibody-binding pattern,altered isoelectric point, direct amino acid sequencing, and any otherof the known assay techniques useful for detecting mutations in aprotein. Such an assay can be provided in a single detection format or amulti-detection format such as an antibody chip array.

[0084] In vitro techniques for detection of peptide include enzymelinked immunosorbent assays (ELISAs), Western blots,immunoprecipitations and immunofluorescence using a detection reagent,such as an antibody or protein binding agent. Alternatively, the peptidecan be detected in vivo in a subject by introducing into the subject alabeled anti-peptide antibody or other types of detection agent. Forexample, the antibody can be labeled with a radioactive marker whosepresence and location in a subject can be detected by standard imagingtechniques. Particularly useful are methods that detect the allelicvariant of a peptide expressed in a subject and methods which detectfragments of a peptide in a sample.

[0085] The peptides are also useful in pharmacogenomic analysis.Pharmacogenomics deal with clinically significant hereditary variationsin the response to drugs due to altered drug disposition and abnormalaction in affected persons. See, e.g., Eichelbaum, M. (Clin. Exp.Pharmacol. Physiol. 23(10-11):983-985 (1996)), and Linder, M. W. (Clin.Chem. 43(2):254-266 (1997)). The clinical outcomes of these variationsresult in severe toxicity of therapeutic drugs in certain individuals ortherapeutic failure of drugs in certain individuals as a result ofindividual variation in metabolism. Thus, the genotype of the individualcan determine the way a therapeutic compound acts on the body or the waythe body metabolizes the compound. Further, the activity of drugmetabolizing enzymes effects both the intensity and duration of drugaction. Thus, the pharmacogenomics of the individual permit theselection of effective compounds and effective dosages of such compoundsfor prophylactic or therapeutic treatment based on the individual'sgenotype. The discovery of genetic polymorphisms in some drugmetabolizing enzymes has explained why some patients do not obtain theexpected drug effects, show an exaggerated drug effect, or experienceserious toxicity from standard drug dosages. Polymorphisms can beexpressed in the phenotype of the extensive metabolizer and thephenotype of the poor metabolizer. Accordingly, genetic polymorphism maylead to allelic protein variants of the kinase protein in which one ormore of the kinase functions in one population is different from thosein another population. The peptides thus allow a target to ascertain agenetic predisposition that can affect treatment modality. Thus, in aligand-based treatment, polymorphism may give rise to amino terminalextracellular domains and/or other substrate-binding regions that aremore or less active in substrate binding, and kinase activation.Accordingly, substrate dosage would necessarily be modified to maximizethe therapeutic effect within a given population containing apolymorphism. As an alternative to genotyping, specific polymorphicpeptides could be identified.

[0086] The peptides are also useful for treating a disordercharacterized by an absence of, inappropriate, or unwanted expression ofthe protein. Experimental data as provided in FIG. 1 indicatesexpression in the human placenta, kidney, lung, skeletal muscle, heart,fetal brain, and colon carcinoma. Accordingly, methods for treatmentinclude the use of the kinase protein or fragments.

[0087] Antibodies

[0088] The invention also provides antibodies that selectively bind toone of the peptides of the present invention, a protein comprising sucha peptide, as well as variants and fragments thereof. As used herein, anantibody selectively binds a target peptide when it binds the targetpeptide and does not significantly bind to unrelated proteins. Anantibody is still considered to selectively bind a peptide even if italso binds to other proteins that are not substantially homologous withthe target peptide so long as such proteins share homology with afragment or domain of the peptide target of the antibody. In this case,it would be understood that antibody binding to the peptide is stillselective despite some degree of cross-reactivity.

[0089] As used herein, an antibody is defined in terms consistent withthat recognized within the art: they are multi-subunit proteins producedby a mammalian organism in response to an antigen challenge. Theantibodies of the present invention include polyclonal antibodies andmonoclonal antibodies, as well as fragments of such antibodies,including, but not limited to, Fab or F(ab′)2, and Fv fragments.

[0090] Many methods are known for generating and/or identifyingantibodies to a given target peptide. Several such methods are describedby Harlow, Antibodies, Cold Spring Harbor Press, (1989).

[0091] In general, to generate antibodies, an isolated peptide is usedas an immunogen and is administered to a mammalian organism, such as arat, rabbit or mouse. The full-length protein, an antigenic peptidefragment or a fusion protein can be used. Particularly importantfragments are those covering functional domains, such as the domainsidentified in FIG. 2, and domain of sequence homology or divergenceamongst the family, such as those that can readily be identified usingprotein alignment methods and as presented in the Figures.

[0092] Antibodies are preferably prepared from regions or discretefragments of the kinase proteins. Antibodies can be prepared from anyregion of the peptide as described herein. However, preferred regionswill include those involved in function/activity and/or kinase/bindingpartner interaction. FIG. 2 can be used to identify particularlyimportant regions while sequence alignment can be used to identifyconserved and unique sequence fragments.

[0093] An antigenic fragment will typically comprise at least 8contiguous amino acid residues. The antigenic peptide can comprise,however, at least 10, 12, 14, 16 or more amino acid residues. Suchfragments can be selected on a physical property, such as fragmentscorrespond to regions that are located on the surface of the protein,e.g., hydrophilic regions or can be selected based on sequenceuniqueness (see FIG. 2).

[0094] Detection on an antibody of the present invention can befacilitated by coupling (i.e., physically linking) the antibody to adetectable substance. Examples of detectable substances include variousenzymes, prosthetic groups, fluorescent materials, luminescentmaterials, bioluminescent materials, and radioactive materials. Examplesof suitable enzymes include horseradish peroxidase, alkalinephosphatase, P-galactosidase, or acetylcholinesterase; examples ofsuitable prosthetic group complexes include streptavidin/biotin andavidin/biotin; examples of suitable fluorescent materials includeumbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine,dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; anexample of a luminescent material includes luminol; examples ofbioluminescent materials include luciferase, luciferin, and aequorin,and examples of suitable radioactive material include ¹²⁵I, ¹³¹I, ³⁵S or³H.

[0095] Antibody Uses

[0096] The antibodies can be used to isolate one of the proteins of thepresent invention by standard techniques, such as affinitychromatography or immunoprecipitation. The antibodies can facilitate thepurification of the natural protein from cells and recombinantlyproduced protein expressed in host cells. In addition, such antibodiesare useful to detect the presence of one of the proteins of the presentinvention in cells or tissues to determine the pattern of expression ofthe protein among various tissues in an organism and over the course ofnormal development. Experimental data as provided in FIG. 1 indicatesthat kinase proteins of the present invention are expressed in the humanplacenta, kidney, lung, skeletal muscle, heart, fetal brain, and coloncarcinoma. Specifically, a virtual northern blot shows expression inhuman colon carcinoma. In addition, PCR-based tissue screening panelindicates expression in human placenta, kidney, lung, skeletal muscle,heart, and fetal brain. Further, such antibodies can be used to detectprotein in situ, in vitro, or in a cell lysate or supernatant in orderto evaluate the abundance and pattern of expression. Also, suchantibodies can be used to assess abnormal tissue distribution orabnormal expression during development or progression of a biologicalcondition. Antibody detection of circulating fragments of the fulllength protein can be used to identify turnover.

[0097] Further, the antibodies can be used to assess expression indisease states such as in active stages of the disease or in anindividual with a predisposition toward disease related to the protein'sfunction. When a disorder is caused by an inappropriate tissuedistribution, developmental expression, level of expression of theprotein, or expressed/processed form, the antibody can be preparedagainst the normal protein. Experimental data as provided in FIG. 1indicates expression in the human placenta, kidney, lung, skeletalmuscle, heart, fetal brain, and colon carcinoma. If a disorder ischaracterized by a specific mutation in the protein, antibodies specificfor this mutant protein can be used to assay for the presence of thespecific mutant protein.

[0098] The antibodies can also be used to assess normal and aberrantsubcellular localization of cells in the various tissues in an organism.Experimental data as provided in FIG. 1 indicates expression in thehuman placenta, kidney, lung, skeletal muscle, heart, fetal brain, andcolon carcinoma. The diagnostic uses can be applied, not only in genetictesting, but also in monitoring a treatment modality. Accordingly, wheretreatment is ultimately aimed at correcting expression level or thepresence of aberrant sequence and aberrant tissue distribution ordevelopmental expression, antibodies directed against the protein orrelevant fragments can be used to monitor therapeutic efficacy.

[0099] Additionally, antibodies are useful in pharmacogenomic analysis.Thus, antibodies prepared against polymorphic proteins can be used toidentify individuals that require modified treatment modalities. Theantibodies are also useful as diagnostic tools as an immunologicalmarker for aberrant protein analyzed by electrophoretic mobility,isoelectric point, tryptic peptide digest, and other physical assaysknown to those in the art.

[0100] The antibodies are also useful for tissue typing. Experimentaldata as provided in FIG. 1 indicates expression in the human placenta,kidney, lung, skeletal muscle, heart, fetal brain, and colon carcinoma.Thus, where a specific protein has been correlated with expression in aspecific tissue, antibodies that are specific for this protein can beused to identify a tissue type.

[0101] The antibodies are also useful for inhibiting protein function,for example, blocking the binding of the kinase peptide to a bindingpartner such as a substrate. These uses can also be applied in atherapeutic context in which treatment involves inhibiting the protein'sfunction. An antibody can be used, for example, to block binding, thusmodulating (agonizing or antagonizing) the peptides activity. Antibodiescan be prepared against specific fragments containing sites required forfunction or against intact protein that is associated with a cell orcell membrane. See FIG. 2 for structural information relating to theproteins of the present invention.

[0102] The invention also encompasses kits for using antibodies todetect the presence of a protein in a biological sample. The kit cancomprise antibodies such as a labeled or labelable antibody and acompound or agent for detecting protein in a biological sample; meansfor determining the amount of protein in the sample; means for comparingthe amount of protein in the sample with a standard; and instructionsfor use. Such a kit can be supplied to detect a single protein orepitope or can be configured to detect one of a multitude of epitopes,such as in an antibody detection array. Arrays are described in detailbelow for nuleic acid arrays and similar methods have been developed forantibody arrays.

[0103] Nucleic Acid Molecules

[0104] The present invention further provides isolated nucleic acidmolecules that encode a kinase peptide or protein of the presentinvention (cDNA, transcript and genomic sequence). Such nucleic acidmolecules will consist of, consist essentially of, or comprise anucleotide sequence that encodes one of the kinase peptides of thepresent invention, an allelic variant thereof, or an ortholog or paralogthereof.

[0105] As used herein, an “isolated” nucleic acid molecule is one thatis separated from other nucleic acid present in the natural source ofthe nucleic acid. Preferably, an “isolated” nucleic acid is free ofsequences which naturally flank the nucleic acid (i.e., sequenceslocated at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA ofthe organism from which the nucleic acid is derived. However, there canbe some flanking nucleotide sequences, for example up to about 5 KB, 4KB, 3 KB, 2 KB, or 1 KB or less, particularly contiguous peptideencoding sequences and peptide encoding sequences within the same genebut separated by introns in the genomic sequence. The important point isthat the nucleic acid is isolated from remote and unimportant flankingsequences such that it can be subjected to the specific manipulationsdescribed herein such as recombinant expression, preparation of probesand primers, and other uses specific to the nucleic acid sequences.

[0106] Moreover, an “isolated” nucleic acid molecule, such as atranscript/cDNA molecule, can be substantially free of other cellularmaterial, or culture medium when produced by recombinant techniques, orchemical precursors or other chemicals when chemically synthesized.However, the nucleic acid molecule can be fused to other coding orregulatory sequences and still be considered isolated.

[0107] For example, recombinant DNA molecules contained in a vector areconsidered isolated. Further examples of isolated DNA molecules includerecombinant DNA molecules maintained in heterologous host cells orpurified (partially or substantially) DNA molecules in solution.Isolated RNA molecules include in vivo or in vitro RNA transcripts ofthe isolated DNA molecules of the present invention. Isolated nucleicacid molecules according to the present invention further include suchmolecules produced synthetically.

[0108] Accordingly, the present invention provides nucleic acidmolecules that consist of the nucleotide sequence shown in FIG. 1 or 3(SEQ ID NO: 1, transcript sequence and SEQ ID NO:3, genomic sequence),or any nucleic acid molecule that encodes the protein provided in FIG.2, SEQ ID NO:2. A nucleic acid molecule consists of a nucleotidesequence when the nucleotide sequence is the complete nucleotidesequence of the nucleic acid molecule.

[0109] The present invention further provides nucleic acid moleculesthat consist essentially of the nucleotide sequence shown in FIG. 1 or 3(SEQ ID NO: 1, transcript sequence and SEQ ID NO:3, genomic sequence),or any nucleic acid molecule that encodes the protein provided in FIG.2, SEQ ID NO:2. A nucleic acid molecule consists essentially of anucleotide sequence when such a nucleotide sequence is present with onlya few additional nucleic acid residues in the final nucleic acidmolecule.

[0110] The present invention further provides nucleic acid moleculesthat comprise the nucleotide sequences shown in FIG. 1 or 3 (SEQ IDNO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or anynucleic acid molecule that encodes the protein provided in FIG. 2, SEQID NO:2. A nucleic acid molecule comprises a nucleotide sequence whenthe nucleotide sequence is at least part of the final nucleotidesequence of the nucleic acid molecule. In such a fashion, the nucleicacid molecule can be only the nucleotide sequence or have additionalnucleic acid residues, such as nucleic acid residues that are naturallyassociated with it or heterologous nucleotide sequences. Such a nucleicacid molecule can have a few additional nucleotides or can comprisesseveral hundred or more additional nucleotides. A brief description ofhow various types of these nucleic acid molecules can be readilymade/isolated is provided below.

[0111] In FIGS. 1 and 3, both coding and non-coding sequences areprovided. Because of the source of the present invention, humans genomicsequence (FIG. 3) and cDNA/transcript sequences (FIG. 1), the nucleicacid molecules in the Figures will contain genomic intronic sequences,5′ and 3′ non-coding sequences, gene regulatory regions and non-codingintergenic sequences. In general such sequence features are either notedin FIGS. 1 and 3 or can readily be identified using computational toolsknown in the art. As discussed below, some of the non-coding regions,particularly gene regulatory elements such as promoters, are useful fora variety of purposes, e.g. control of heterologous gene expression,target for identifying gene activity modulating compounds, and areparticularly claimed as fragments of the genomic sequence providedherein.

[0112] The isolated nucleic acid molecules can encode the mature proteinplus additional amino or carboxyl-terminal amino acids, or amino acidsinterior to the mature peptide (when the mature form has more than onepeptide chain, for instance). Such sequences may play a role inprocessing of a protein from precursor to a mature form, facilitateprotein trafficking, prolong or shorten protein half-life or facilitatemanipulation of a protein for assay or production, among other things.As generally is the case in situ, the additional amino acids may beprocessed away from the mature protein by cellular enzymes.

[0113] As mentioned above, the isolated nucleic acid molecules include,but are not limited to, the sequence encoding the kinase peptide alone,the sequence encoding the mature peptide and additional codingsequences, such as a leader or secretory sequence (e.g., a pre-pro orpro-protein sequence), the sequence encoding the mature peptide, with orwithout the additional coding sequences, plus additional non-codingsequences, for example introns and non-coding 5′ and 3′ sequences suchas transcribed but non-translated sequences that play a role intranscription, mRNA processing (including splicing and polyadenylationsignals), ribosome binding and stability of mRNA. In addition, thenucleic acid molecule may be fused to a marker sequence encoding, forexample, a peptide that facilitates purification.

[0114] Isolated nucleic acid molecules can be in the form of RNA, suchas mRNA, or in the form DNA, including cDNA and genomic DNA obtained bycloning or produced by chemical synthetic techniques or by a combinationthereof. The nucleic acid, especially DNA, can be double-stranded orsingle-stranded. Single-stranded nucleic acid can be the coding strand(sense strand) or the non-coding strand (anti-sense strand).

[0115] The invention further provides nucleic acid molecules that encodefragments of the peptides of the present invention as well as nucleicacid molecules that encode obvious variants of the kinase proteins ofthe present invention that are described above. Such nucleic acidmolecules may be naturally occurring, such as allelic variants (samelocus), paralogs (different locus), and orthologs (different organism),or may be constructed by recombinant DNA methods or by chemicalsynthesis. Such non-naturally occurring variants may be made bymutagenesis techniques, including those applied to nucleic acidmolecules, cells, or organisms. Accordingly, as discussed above, thevariants can contain nucleotide substitutions, deletions, inversions andinsertions. Variation can occur in either or both the coding andnon-coding regions. The variations can produce both conservative andnon-conservative amino acid substitutions.

[0116] The present invention further provides non-coding fragments ofthe nucleic acid molecules provided in FIGS. 1 and 3. Preferrednon-coding fragments include, but are not limited to, promotersequences, enhancer sequences, gene modulating sequences and genetermination sequences. Such fragments are useful in controllingheterologous gene expression and in developing screens to identifygene-modulating agents. A promoter can readily be identified as being 5′to the ATG start site in the genomic sequence provided in FIG. 3.

[0117] A fragment comprises a contiguous nucleotide sequence greaterthan 12 or more nucleotides. Further, a fragment could at least 30, 40,50, 100, 250 or 500 nucleotides in length. The length of the fragmentwill be based on its intended use. For example, the fragment can encodeepitope bearing regions of the peptide, or can be useful as DNA probesand primers. Such fragments can be isolated using the known nucleotidesequence to synthesize an oligonucleotide probe. A labeled probe canthen be used to screen a cDNA library, genomic DNA library, or mRNA toisolate nucleic acid corresponding to the coding region. Further,primers can be used in PCR reactions to clone specific regions of gene.

[0118] A probe/primer typically comprises substantially a purifiedoligonucleotide or oligonucleotide pair. The oligonucleotide typicallycomprises a region of nucleotide sequence that hybridizes understringent conditions to at least about 12, 20, 25, 40, 50 or moreconsecutive nucleotides.

[0119] Orthologs, homologs, and allelic variants can be identified usingmethods well known in the art. As described in the Peptide Section,these variants comprise a nucleotide sequence encoding a peptide that istypically 60-70%, 70-80%, 80-90%, and more typically at least about90-95% or more homologous to the nucleotide sequence shown in the Figuresheets or a fragment of this sequence. Such nucleic acid molecules canreadily be identified as being able to hybridize under moderate tostringent conditions, to the nucleotide sequence shown in the Figuresheets or a fragment of the sequence. Allelic variants can readily bedetermined by genetic locus of the encoding gene. As indicated by thedata presented in FIG. 3, the map position was determined to be onchromosome 1 by ePCR, and confirmed with radiation hybrid mapping. Asindicated by the data presented in FIG. 3, the gene provided by thepresent invention encoding a novel phosphatase maps to public BAC ACAC023889, which is known to be located on human chromosome 1.

[0120]FIG. 3 provides information on SNPs that have been identified in agene encoding the kinase protein of the present invention. 6 SNPvariants were found, and all SNPs in exons, of which 3 of these causechanges in the amino acid sequence (i.e., nonsynonymous SNPs). Thechanges in the amino acid sequence that these SNPs cause is indicated inFIG. 3 and can readily be determined using the universal genetic codeand the protein sequence provided in FIG. 2 as a reference.

[0121] As used herein, the term “hybridizes under stringent conditions”is intended to describe conditions for hybridization and washing underwhich nucleotide sequences encoding a peptide at least 60-70% homologousto each other typically remain hybridized to each other. The conditionscan be such that sequences at least about 60%, at least about 70%, or atleast about 80% or more homologous to each other typically remainhybridized to each other. Such stringent conditions are known to thoseskilled in the art and can be found in Current Protocols in MolecularBiology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. One example ofstringent hybridization conditions are hybridization in 6× sodiumchloride/sodium citrate (SSC) at about 45C, followed by one or morewashes in 0.2×SSC, 0.1% SDS at 50-65C. Examples of moderate to lowstringency hybridization conditions are well known in the art.

[0122] Nucleic Acid Molecule Uses

[0123] The nucleic acid molecules of the present invention are usefulfor probes, primers, chemical intermediates, and in biological assays.The nucleic acid molecules are useful as a hybridization probe formessenger RNA, transcript/cDNA and genomic DNA to isolate full-lengthcDNA and genomic clones encoding the peptide described in FIG. 2 and toisolate cDNA and genomic clones that correspond to variants (alleles,orthologs, etc.) producing the same or related peptides shown in FIG. 2.6 SNPs, have been identified in the gene encoding the kinase proteinprovided by the present invention and are given in FIG. 3.

[0124] The probe can correspond to any sequence along the entire lengthof the nucleic acid molecules provided in the Figures. Accordingly, itcould be derived from 5′ noncoding regions, the coding region, and 3′noncoding regions. However, as discussed, fragments are not to beconstrued as encompassing fragments disclosed prior to the presentinvention.

[0125] The nucleic acid molecules are also useful as primers for PCR toamplify any given region of a nucleic acid molecule and are useful tosynthesize antisense molecules of desired length and sequence.

[0126] The nucleic acid molecules are also useful for constructingrecombinant vectors. Such vectors include expression vectors thatexpress a portion of, or all of, the peptide sequences. Vectors alsoinclude insertion vectors, used to integrate into another nucleic acidmolecule sequence, such as into the cellular genome, to alter in situexpression of a gene and/or gene product. For example, an endogenouscoding sequence can be replaced via homologous recombination with all orpart of the coding region containing one or more specifically introducedmutations.

[0127] The nucleic acid molecules are also useful for expressingantigenic portions of the proteins.

[0128] The nucleic acid molecules are also useful as probes fordetermining the chromosomal positions of the nucleic acid molecules bymeans of in situ hybridization methods. As indicated by the datapresented in FIG. 3, the map position was determined to be on chromosome1 by ePCR, and confirmed with radiation hybrid mapping. As indicated bythe data presented in FIG. 3, the gene provided by the present inventionencoding a novel phosphatase maps to public BAC AC AC023889, which isknown to be located on human chromosome 1.

[0129] The nucleic acid molecules are also useful in making vectorscontaining the gene regulatory regions of the nucleic acid molecules ofthe present invention.

[0130] The nucleic acid molecules are also useful for designingribozymes corresponding to all, or a part, of the mRNA produced from thenucleic acid molecules described herein.

[0131] The nucleic acid molecules are also useful for making vectorsthat express part, or all, of the peptides.

[0132] The nucleic acid molecules are also useful for constructing hostcells expressing a part, or all, of the nucleic acid molecules andpeptides.

[0133] The nucleic acid molecules are also useful for constructingtransgenic animals expressing ail, or a part, of the nucleic acidmolecules and peptides.

[0134] The nucleic acid molecules are also useful as hybridizationprobes for determining the presence, level, form and distribution ofnucleic acid expression. Experimental data as provided in FIG. 1indicates that kinase proteins of the present invention are expressed inthe human placenta, kidney, lung, skeletal muscle, heart, fetal brain,and colon carcinoma. Specifically, a virtual northern blot showsexpression in human colon carcinoma. In addition, PCR-based tissuescreening panel indicates expression in human placenta, kidney, lung,skeletal muscle, heart, and fetal brain. Accordingly, the probes can beused to detect the presence of, or to determine levels of, a specificnucleic acid molecule in cells, tissues, and in organisms. The nucleicacid whose level is determined can be DNA or RNA. Accordingly, probescorresponding to the peptides described herein can be used to assessexpression and/or gene copy number in a given cell, tissue, or organism.These uses are relevant for diagnosis of disorders involving an increaseor decrease in kinase protein expression relative to normal results.

[0135] In vitro techniques for detection of mRNA include Northernhybridizations and in situ hybridizations. In vitro techniques fordetecting DNA includes Southern hybridizations and in situhybridization.

[0136] Probes can be used as a part of a diagnostic test kit foridentifying cells or tissues that express a kinase protein, such as bymeasuring a level of a kinase-encoding nucleic acid in a sample of cellsfrom a subject e.g., mRNA or genomic DNA, or determining if a kinasegene has been mutated. Experimental data as provided in FIG. 1 indicatesthat kinase proteins of the present invention are expressed in the humanplacenta, kidney, lung, skeletal muscle, heart, fetal brain, and coloncarcinoma. Specifically, a virtual northern blot shows expression inhuman colon carcinoma. In addition, PCR-based tissue screening panelindicates expression in human placenta, kidney, lung, skeletal muscle,heart, and fetal brain.

[0137] Nucleic acid expression assays are useful for drug screening toidentify compounds that modulate kinase nucleic acid expression.

[0138] The invention thus provides a method for identifying a compoundthat can be used to treat a disorder associated with nucleic acidexpression of the kinase gene, particularly biological and pathologicalprocesses that are mediated by the kinase in cells and tissues thatexpress it. Experimental data as provided in FIG. 1 indicates expressionin the human placenta, kidney, lung, skeletal muscle, heart, fetalbrain, and colon carcinoma. The method typically includes assaying theability of the compound to modulate the expression of the kinase nucleicacid and thus identifying a compound that can be used to treat adisorder characterized by undesired kinase nucleic acid expression. Theassays can be performed in cell-based and cell-free systems. Cell-basedassays include cells naturally expressing the kinase nucleic acid orrecombinant cells genetically engineered to express specific nucleicacid sequences.

[0139] The assay for kinase nucleic acid expression can involve directassay of nucleic acid levels, such as mRNA levels, or on collateralcompounds involved in the signal pathway. Further, the expression ofgenes that are up- or down-regulated in response to the kinase proteinsignal pathway can also be assayed. In this embodiment the regulatoryregions of these genes can be operably linked to a reporter gene such asluciferase.

[0140] Thus, modulators of kinase gene expression can be identified in amethod wherein a cell is contacted with a candidate compound and theexpression of mRNA determined. The level of expression of kinase mRNA inthe presence of the candidate compound is compared to the level ofexpression of kinase mRNA in the absence of the candidate compound. Thecandidate compound can then be identified as a modulator of nucleic acidexpression based on this comparison and be used, for example to treat adisorder characterized by aberrant nucleic acid expression. Whenexpression of mRNA is statistically significantly greater in thepresence of the candidate compound than in its absence, the candidatecompound is identified as a stimulator of nucleic acid expression. Whennucleic acid expression is statistically significantly less in thepresence of the candidate compound than in its absence, the candidatecompound is identified as an inhibitor of nucleic acid expression.

[0141] The invention further provides methods of treatment, with thenucleic acid as a target, using a compound identified through drugscreening as a gene modulator to modulate kinase nucleic acid expressionin cells and tissues that express the kinase. Experimental data asprovided in FIG. 1 indicates that kinase proteins of the presentinvention are expressed in the human placenta, kidney, lung, skeletalmuscle, heart, fetal brain, and colon carcinoma. Specifically, a virtualnorthern blot shows expression in human colon carcinoma. In addition,PCR-based tissue screening panel indicates expression in human placenta,kidney, lung, skeletal muscle, heart, and fetal brain. Modulationincludes both up-regulation (i.e. activation or agonization) ordown-regulation (suppression or antagonization) or nucleic acidexpression.

[0142] Alternatively, a modulator for kinase nucleic acid expression canbe a small molecule or drug identified using the screening assaysdescribed herein as long as the drug or small molecule inhibits thekinase nucleic acid expression in the cells and tissues that express theprotein. Experimental data as provided in FIG. 1 indicates expression inthe human placenta, kidney, lung, skeletal muscle, heart, fetal brain,and colon carcinoma.

[0143] The nucleic acid molecules are also useful for monitoring theeffectiveness of modulating compounds on the expression or activity ofthe kinase gene in clinical trials or in a treatment regimen. Thus, thegene expression pattern can serve as a barometer for the continuingeffectiveness of treatment with the compound, particularly withcompounds to which a patient can develop resistance. The gene expressionpattern can also serve as a marker indicative of a physiologicalresponse of the affected cells to the compound. Accordingly, suchmonitoring would allow either increased administration of the compoundor the administration of alternative compounds to which the patient hasnot become resistant. Similarly, if the level of nucleic acid expressionfalls below a desirable level, administration of the compound could becommensurately decreased.

[0144] The nucleic acid molecules are also useful in diagnostic assaysfor qualitative changes in kinase nucleic acid expression, andparticularly in qualitative changes that lead to pathology. The nucleicacid molecules can be used to detect mutations in kinase genes and geneexpression products such as mRNA. The nucleic acid molecules can be usedas hybridization probes to detect naturally occurring genetic mutationsin the kinase gene and thereby to determine whether a subject with themutation is at risk for a disorder caused by the mutation. Mutationsinclude deletion, addition, or substitution of one or more nucleotidesin the gene, chromosomal rearrangement, such as inversion ortransposition, modification of genomic DNA, such as aberrant methylationpatterns or changes in gene copy number, such as amplification.Detection of a mutated form of the kinase gene associated with adysfunction provides a diagnostic tool for an active disease orsusceptibility to disease when the disease results from overexpression,underexpression, or altered expression of a kinase protein.

[0145] Individuals carrying mutations in the kinase gene can be detectedat the nucleic acid level by a variety of techniques. FIG. 3 providesinformation on SNPs that have been identified in a gene encoding thekinase protein of the present invention. 6 SNP variants were found, andall SNPs in exons, of which 3 of these cause changes in the amino acidsequence (i.e., nonsynonymous SNPs). The changes in the amino acidsequence that these SNPs cause is indicated in FIG. 3 and can readily bedetermined using the universal genetic code and the protein sequenceprovided in FIG. 2 as a reference. As indicated by the data presented inFIG. 3, the map position was determined to be on chromosome 1 by ePCR,and confirmed with radiation hybrid mapping. As indicated by the datapresented in FIG. 3, the gene provided by the present invention encodinga novel phosphatase maps to public BAC AC AC023889, which is known to belocated on human chromosome 1. Genomic DNA can be analyzed directly orcan be amplified by using PCR prior to analysis. RNA or cDNA can be usedin the same way. In some uses, detection of the mutation involves theuse of a probe/primer in a polymerase chain reaction (PCR) (see, e.g.U.S. Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR,or, alternatively, in a ligation chain reaction (LCR) (see, e.g.,Landegran et al., Science 241:1077-1080 (1988); and Nakazawa et al.,PNAS 91:360-364 (1994)), the latter of which can be particularly usefulfor detecting point mutations in the gene (see Abravaya et al., NucleicAcids Res. 23:675-682 (1995)). This method can include the steps ofcollecting a sample of cells from a patient, isolating nucleic acid(e.g., genomic, mRNA or both) from the cells of the sample, contactingthe nucleic acid sample with one or more primers which specificallyhybridize to a gene under conditions such that hybridization andamplification of the gene (if present) occurs, and detecting thepresence or absence of an amplification product, or detecting the sizeof the amplification product and comparing the length to a controlsample. Deletions and insertions can be detected by a change in size ofthe amplified product compared to the normal genotype. Point mutationscan be identified by hybridizing amplified DNA to normal RNA orantisense DNA sequences.

[0146] Alternatively, mutations in a kinase gene can be directlyidentified, for example, by alterations in restriction enzyme digestionpatterns determined by gel electrophoresis.

[0147] Further, sequence-specific ribozymes (U.S. Pat. No. 5,498,531)can be used to score for the presence of specific mutations bydevelopment or loss of a ribozyme cleavage site. Perfectly matchedsequences can be distinguished from mismatched sequences by nucleasecleavage digestion assays or by differences in melting temperature.

[0148] Sequence changes at specific locations can also be assessed bynuclease protection assays such as RNase and S1 protection or thechemical cleavage method. Furthermore, sequence differences between amutant kinase gene and a wild-type gene can be determined by direct DNAsequencing. A variety of automated sequencing procedures can be utilizedwhen performing the diagnostic assays (Naeve, C. W., (1995)Biotechniques 19:448), including sequencing by mass spectrometry (see,e.g., PCT International Publication No. WO 94/16101; Cohen et al., Adv.Chromatogr. 36:127-162 (1996); and Griffin et al., Appl. Biochem.Biotechnol. 38:147-159 (1993)).

[0149] Other methods for detecting mutations in the gene include methodsin which protection from cleavage agents is used to detect mismatchedbases in RNA/RNA or RNA/DNA duplexes (Myers et al., Science 230:1242(1985)); Cotton et al., PNAS 85:4397 (1988); Saleeba et al., Meth.EnzymoL 217:286-295 (1992)), electrophoretic mobility of mutant and wildtype nucleic acid is compared (Orita et al., PNAS 86:2766 (1989); Cottonet al., Mutat. Res. 285:125-144 (1993); and Hayashi et al., Genet. Anal.Tech. Appl. 9:73-79 (1992)), and movement of mutant or wild-typefragments in polyacrylamide gels containing a gradient of denaturant isassayed using denaturing gradient gel electrophoresis (Myers et al.,Nature 313:495 (1985)). Examples of other techniques for detecting pointmutations include selective oligonucleotide hybridization, selectiveamplification, and selective primer extension.

[0150] The nucleic acid molecules are also useful for testing anindividual for a genotype that while not necessarily causing thedisease, nevertheless affects the treatment modality. Thus, the nucleicacid molecules can be used to study the relationship between anindividual's genotype and the individual's response to a compound usedfor treatment (pharmacogenomic relationship). Accordingly, the nucleicacid molecules described herein can be used to assess the mutationcontent of the kinase gene in an individual in order to select anappropriate compound or dosage regimen for treatment. FIG. 3 providesinformation on SNPs that have been identified in a gene encoding thekinase protein of the present invention. 6 SNP variants were found, andall SNPs in exons, of which 3 of these cause changes in the amino acidsequence (i.e., nonsynonymous SNPs). The changes in the amino acidsequence that these SNPs cause is indicated in FIG. 3 and can readily bedetermined using the universal genetic code and the protein sequenceprovided in FIG. 2 as a reference.

[0151] Thus nucleic acid molecules displaying genetic variations thataffect treatment provide a diagnostic target that can be used to tailortreatment in an individual. Accordingly, the production of recombinantcells and animals containing these polymorphisms allow effectiveclinical design of treatment compounds and dosage regimens.

[0152] The nucleic acid molecules are thus useful as antisenseconstructs to control kinase gene expression in cells, tissues, andorganisms. A DNA antisense nucleic acid molecule is designed to becomplementary to a region of the gene involved in transcription,preventing transcription and hence production of kinase protein. Anantisense RNA or DNA nucleic acid molecule would hybridize to the mRNAand thus block translation of mRNA into kinase protein.

[0153] Alternatively, a class of antisense molecules can be used toinactivate mRNA in order to decrease expression of kinase nucleic acid.Accordingly, these molecules can treat a disorder characterized byabnormal or undesired kinase nucleic acid expression. This techniqueinvolves cleavage by means of ribozymes containing nucleotide sequencescomplementary to one or more regions in the mRNA that attenuate theability of the mRNA to be translated. Possible regions include codingregions and particularly coding regions corresponding to the catalyticand other functional activities of the kinase protein, such as substratebinding.

[0154] The nucleic acid molecules also provide vectors for gene therapyin patients containing cells that are aberrant in kinase geneexpression. Thus, recombinant cells, which include the patient's cellsthat have been engineered ex vivo and returned to the patient, areintroduced into an individual where the cells produce the desired kinaseprotein to treat the individual.

[0155] The invention also encompasses kits for detecting the presence ofa kinase nucleic acid in a biological sample. Experimental data asprovided in FIG. 1 indicates that kinase proteins of the presentinvention are expressed in the human placenta, kidney, lung, skeletalmuscle, heart, fetal brain, and colon carcinoma. Specifically, a virtualnorthern blot shows expression in human colon carcinoma. In addition,PCR-based tissue screening panel indicates expression in human placenta,kidney, lung, skeletal muscle, heart, and fetal brain. For example, thekit can comprise reagents such as a labeled or labelable nucleic acid oragent capable of detecting kinase nucleic acid in a biological sample;means for determining the amount of kinase nucleic acid in the sample;and means for comparing the amount of kinase nucleic acid in the samplewith a standard. The compound or agent can be packaged in a suitablecontainer. The kit can further comprise instructions for using the kitto detect kinase protein mRNA or DNA.

[0156] Nucleic Acid Arrays

[0157] The present invention further provides nucleic acid detectionkits, such as arrays or microarrays of nucleic acid molecules that arebased on the sequence information provided in FIGS. 1 and 3 (SEQ IDNOS:1 and 3).

[0158] As used herein “Arrays” or “Microarrays” refers to an array ofdistinct polynucleotides or oligonucleotides synthesized on a substrate,such as paper, nylon or other type of membrane, filter, chip, glassslide, or any other suitable solid support. In one embodiment, themicroarray is prepared and used according to the methods described inU.S. Pat. No. 5,837,832, Chee et al., PCT application WO95/11995 (Cheeet al.), Lockhart, D. J. et al. (1996; Nat. Biotech. 14: 1675-1680) andSchena, M. et al. (1996; Proc. Natl. Acad. Sci. 93: 10614-10619), all ofwhich are incorporated herein in their entirety by reference. In otherembodiments, such arrays are produced by the methods described by Brownet al., U.S. Pat. No. 5,807,522.

[0159] The microarray or detection kit is preferably composed of a largenumber of unique, single-stranded nucleic acid sequences, usually eithersynthetic antisense oligonucleotides or fragments of cDNAs, fixed to asolid support. The oligonucleotides are preferably about 6-60nucleotides in length, more preferably 15-30 nucleotides in length, andmost preferably about 20-25 nucleotides in length. For a certain type ofmicroarray or detection kit, it may be preferable to useoligonucleotides that are only 7-20 nucleotides in length. Themicroarray or detection kit may contain oligonucleotides that cover theknown 5′, or 3′, sequence, sequential oligonucleotides which cover thefull length sequence; or unique oligonucleotides selected fromparticular areas along the length of the sequence. Polynucleotides usedin the microarray or detection kit may be oligonucleotides that arespecific to a gene or genes of interest.

[0160] In order to produce oligonucleotides to a known sequence for amicroarray or detection kit, the gene(s) of interest (or an ORFidentified from the contigs of the present invention) is typicallyexamined using a computer algorithm which starts at the 5′ or at the 3′end of the nucleotide sequence. Typical algorithms will then identifyoligomers of defined length that are unique to the gene, have a GCcontent within a range suitable for hybridization, and lack predictedsecondary structure that may interfere with hybridization. In certainsituations it may be appropriate to use pairs of oligonucleotides on amicroarray or detection kit. The “pairs” will be identical, except forone nucleotide that preferably is located in the center of the sequence.The second oligonucleotide in the pair (mismatched by one) serves as acontrol. The number of oligonucleotide pairs may range from two to onemillion. The oligomers are synthesized at designated areas on asubstrate using a light-directed chemical process. The substrate may bepaper, nylon or other type of membrane, filter, chip, glass slide or anyother suitable solid support.

[0161] In another aspect, an oligonucleotide may be synthesized on thesurface of the substrate by using a chemical coupling procedure and anink jet application apparatus, as described in PCT applicationWO95/251116 (Baldeschweiler et al.) which is incorporated herein in itsentirety by reference. In another aspect, a “gridded” array analogous toa dot (or slot) blot may be used to arrange and link cDNA fragments oroligonucleotides to the surface of a substrate using a vacuum system,thermal, UV, mechanical or chemical bonding procedures. An array, suchas those described above, may be produced by hand or by using availabledevices (slot blot or dot blot apparatus), materials (any suitable solidsupport), and machines (including robotic instruments), and may contain8, 24, 96, 384, 1536, 6144 or more oligonucleotides, or any other numberbetween two and one million which lends itself to the efficient use ofcommercially available instrumentation.

[0162] In order to conduct sample analysis using a microarray ordetection kit, the RNA or DNA from a biological sample is made intohybridization probes. The mRNA is isolated, and cDNA is produced andused as a template to make antisense RNA (aRNA). The aRNA is amplifiedin the presence of fluorescent nucleotides, and labeled probes areincubated with the microarray or detection kit so that the probesequences hybridize to complementary oligonucleotides of the microarrayor detection kit. Incubation conditions are adjusted so thathybridization occurs with precise complementary matches or with variousdegrees of less complementarity. After removal of nonhybridized probes,a scanner is used to determine the levels and patterns of fluorescence.The scanned images are examined to determine degree of complementarityand the relative abundance of each oligonucleotide sequence on themicroarray or detection kit. The biological samples may be obtained fromany bodily fluids (such as blood, urine, saliva, phlegm, gastric juices,etc.), cultured cells, biopsies, or other tissue preparations. Adetection system may be used to measure the absence, presence, andamount of hybridization for all of the distinct sequencessimultaneously. This data may be used for large-scale correlationstudies on the sequences, expression patterns, mutations, variants, orpolymorphisms among samples.

[0163] Using such arrays, the present invention provides methods toidentify the expression of the kinase proteins/peptides of the presentinvention. In detail, such methods comprise incubating a test samplewith one or more nucleic acid molecules and assaying for binding of thenucleic acid molecule with components within the test sample. Suchassays will typically involve arrays comprising many genes, at least oneof which is a gene of the present invention and or alleles of the kinasegene of the present invention. FIG. 3 provides information on SNPs thathave been identified in a gene encoding the kinase protein of thepresent invention. 6 SNP variants were found, and all SNPs in exons, ofwhich 3 of these cause changes in the amino acid sequence (i.e.,nonsynonymous SNPs). The changes in the amino acid sequence that theseSNPs cause is indicated in FIG. 3 and can readily be determined usingthe universal genetic code and the protein sequence provided in FIG. 2as a reference.

[0164] Conditions for incubating a nucleic acid molecule with a testsample vary. Incubation conditions depend on the format employed in theassay, the detection methods employed, and the type and nature of thenucleic acid molecule used in the assay. One skilled in the art willrecognize that any one of the commonly available hybridization,amplification or array assay formats can readily be adapted to employthe novel fragments of the Human genome disclosed herein. Examples ofsuch assays can be found in Chard, T, An Introduction toRadioimmunoassay and Related Techniques, Elsevier Science Publishers,Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques inImmunocytochemistry, Academic Press, Orlando, Fla. Vol. 1 (1982), Vol. 2(1983), Vol. 3 (1985); Tijssen, P., Practice and Theory of EnzymeImmunoassays: Laboratory Techniques in Biochemistry and MolecularBiology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985).

[0165] The test samples of the present invention include cells, proteinor membrane extracts of cells. The test sample used in theabove-described method will vary based on the assay format, nature ofthe detection method and the tissues, cells or extracts used as thesample to be assayed. Methods for preparing nucleic acid extracts or ofcells are well known in the art and can be readily be adapted in orderto obtain a sample that is compatible with the system utilized.

[0166] In another embodiment of the present invention, kits are providedwhich contain the necessary reagents to carry out the assays of thepresent invention.

[0167] Specifically, the invention provides a compartmentalized kit toreceive, in close confinement, one or more containers which comprises:(a) a first container comprising one of the nucleic acid molecules thatcan bind to a fragment of the Human genome disclosed herein; and (b) oneor more other containers comprising one or more of the following: washreagents, reagents capable of detecting presence of a bound nucleicacid.

[0168] In detail, a compartmentalized kit includes any kit in whichreagents are contained in separate containers. Such containers includesmall glass containers, plastic containers, strips of plastic, glass orpaper, or arraying material such as silica. Such containers allows oneto efficiently transfer reagents from one compartment to anothercompartment such that the samples and reagents are notcross-contaminated, and the agents or solutions of each container can beadded in a quantitative fashion from one compartment to another. Suchcontainers will include a container which will accept the test sample, acontainer which contains the nucleic acid probe, containers whichcontain wash reagents (such as phosphate buffered saline, Tris-buffers,etc.), and containers which contain the reagents used to detect thebound probe. One skilled in the art will readily recognize that thepreviously unidentified kinase gene of the present invention can beroutinely identified using the sequence information disclosed herein canbe readily incorporated into one of the established kit formats whichare well known in the art, particularly expression arrays.

[0169] Vectors/Host Cells

[0170] The invention also provides vectors containing the nucleic acidmolecules described herein. The term “vector” refers to a vehicle,preferably a nucleic acid molecule, which can transport the nucleic acidmolecules. When the vector is a nucleic acid molecule, the nucleic acidmolecules are covalently linked to the vector nucleic acid. With thisaspect of the invention, the vector includes a plasmid, single or doublestranded phage, a single or double stranded RNA or DNA viral vector, orartificial chromosome, such as a BAC, PAC, YAC, OR MAC.

[0171] A vector can be maintained in the host cell as anextrachromosomal element where it replicates and produces additionalcopies of the nucleic acid molecules. Alternatively, the vector mayintegrate into the host cell genome and produce additional copies of thenucleic acid molecules when the host cell replicates.

[0172] The invention provides vectors for the maintenance (cloningvectors) or vectors for expression (expression vectors) of the nucleicacid molecules. The vectors can function in prokaryotic or eukaryoticcells or in both (shuttle vectors).

[0173] Expression vectors contain cis-acting regulatory regions that areoperably linked in the vector to the nucleic acid molecules such thattranscription of the nucleic acid molecules is allowed in a host cell.The nucleic acid molecules can be introduced into the host cell with aseparate nucleic acid molecule capable of affecting transcription. Thus,the second nucleic acid molecule may provide a trans-acting factorinteracting with the cis-regulatory control region to allowtranscription of the nucleic acid molecules from the vector.Alternatively, a trans-acting factor may be supplied by the host cell.Finally, a trans-acting factor can be produced from the vector itself.It is understood, however, that in some embodiments, transcriptionand/or translation of the nucleic acid molecules can occur in acell-free system.

[0174] The regulatory sequence to which the nucleic acid moleculesdescribed herein can be operably linked include promoters for directingmRNA transcription. These include, but are not limited to, the leftpromoter from bacteriophage λ, the lac, TRP, and TAC promoters from E.coli, the early and late promoters from SV40, the CMV immediate earlypromoter, the adenovirus early and late promoters, and retroviruslong-terminal repeats.

[0175] In addition to control regions that promote transcription,expression vectors may also include regions that modulate transcription,such as repressor binding sites and enhancers. Examples include the SV40enhancer, the cytomegalovirus immediate early enhancer, polyomaenhancer, adenovirus enhancers, and retrovirus LTR enhancers.

[0176] In addition to containing sites for transcription initiation andcontrol, expression vectors can also contain sequences necessary fortranscription termination and, in the transcribed region a ribosomebinding site for translation. Other regulatory control elements forexpression include initiation and termination codons as well aspolyadenylation signals. The person of ordinary skill in the art wouldbe aware of the numerous regulatory sequences that are useful inexpression vectors. Such regulatory sequences are described, forexample, in Sambrook et al., Molecular Cloning: A Laboratory Manual.2nd. ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.,(1989).

[0177] A variety of expression vectors can be used to express a nucleicacid molecule. Such vectors include chromosomal, episomal, andvirus-derived vectors, for example vectors derived from bacterialplasmids, from bacteriophage, from yeast episomes, from yeastchromosomal elements, including yeast artificial chromosomes, fromviruses such as baculoviruses, papovaviruses such as SV40, Vacciniaviruses, adenoviruses, poxviruses, pseudorabies viruses, andretroviruses. Vectors may also be derived from combinations of thesesources such as those derived from plasmid and bacteriophage geneticelements, e.g. cosmids and phagemids. Appropriate cloning and expressionvectors for prokaryotic and eukaryotic hosts are described in Sambrooket al., Molecular Cloning: A Laboratory Manual. 2nd. ed., Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).

[0178] The regulatory sequence may provide constitutive expression inone or more host cells (i.e. tissue specific) or may provide forinducible expression in one or more cell types such as by temperature,nutrient additive, or exogenous factor such as a hormone or otherligand. A variety of vectors providing for constitutive and inducibleexpression in prokaryotic and eukaryotic hosts are well known to thoseof ordinary skill in the art.

[0179] The nucleic acid molecules can be inserted into the vectornucleic acid by well-known methodology. Generally, the DNA sequence thatwill ultimately be expressed is joined to an expression vector bycleaving the DNA sequence and the expression vector with one or morerestriction enzymes and then ligating the fragments together. Proceduresfor restriction enzyme digestion and ligation are well known to those ofordinary skill in the art.

[0180] The vector containing the appropriate nucleic acid molecule canbe introduced into an appropriate host cell for propagation orexpression using well-known techniques. Bacterial cells include, but arenot limited to, E. coli, Streptomyces, and Salmonella typhimurium.Eukaryotic cells include, but are not limited to, yeast, insect cellssuch as Drosophila, animal cells such as COS and CHO cells, and plantcells.

[0181] As described herein, it may be desirable to express the peptideas a fusion protein. Accordingly, the invention provides fusion vectorsthat allow for the production of the peptides. Fusion vectors canincrease the expression of a recombinant protein, increase thesolubility of the recombinant protein, and aid in the purification ofthe protein by acting for example as a ligand for affinity purification.A proteolytic cleavage site may be introduced at the junction of thefusion moiety so that the desired peptide can ultimately be separatedfrom the fusion moiety. Proteolytic enzymes include, but are not limitedto, factor Xa, thrombin, and enterokinase. Typical fusion expressionvectors include pGEX (Smith et al., Gene 67:31-40 (1988)), pMAL (NewEngland Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.)which fuse glutathione S-transferase (GST), maltose E binding protein,or protein A, respectively, to the target recombinant protein. Examplesof suitable inducible non-fusion E. coli expression vectors include pTrc(Amann et al., Gene 69:301-315 (1988)) and pET 11d (Studier et al., GeneExpression Technology: Methods in Enzymology 185:60-89 (1990)).

[0182] Recombinant protein expression can be maximized in host bacteriaby providing a genetic background wherein the host cell has an impairedcapacity to proteolytically cleave the recombinant protein. (Gottesman,S., Gene Expression Technology: Methods in Enzymology 185, AcademicPress, San Diego, Calif. (1990) 119-128). Alternatively, the sequence ofthe nucleic acid molecule of interest can be altered to providepreferential codon usage for a specific host cell, for example E. coli.(Wada et al., Nucleic Acids Res. 20:2111-2118 (1992)).

[0183] The nucleic acid molecules can also be expressed by expressionvectors that are operative in yeast. Examples of vectors for expressionin yeast e.g., S. cerevisiae include pYepSec1 (Baldari, et al., EMBO J6:229-234 (1987)), pMFa (Kujan et al., Cell 30:933-943(1982)), pJRY88(Schultz et al., Gene 54:113-123 (1987)), and pYES2 (InvitrogenCorporation, San Diego, Calif.).

[0184] The nucleic acid molecules can also be expressed in insect cellsusing, for example, baculovirus expression vectors. Baculovirus vectorsavailable for expression of proteins in cultured insect cells (e.g., Sf9 cells) include the pAc series (Smith et al., Mol. Cell Biol.3:2156-2165 (1983)) and the pVL series (Lucklow et al., Virology170:31-39 (1989)).

[0185] In certain embodiments of the invention, the nucleic acidmolecules described herein are expressed in mammalian cells usingmammalian expression vectors. Examples of mammalian expression vectorsinclude pCDM8 (Seed, B. Nature 329:840(1987)) and pMT2PC (Kaufman etal., EMBO J. 6:187-195 (1987)).

[0186] The expression vectors listed herein are provided by way ofexample only of the well-known vectors available to those of ordinaryskill in the art that would be useful to express the nucleic acidmolecules. The person of ordinary skill in the art would be aware ofother vectors suitable for maintenance propagation or expression of thenucleic acid molecules described herein. These are found for example inSambrook, J., Fritsh, E. F., and Maniatis, T. Molecular Cloning: ALaboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.

[0187] The invention also encompasses vectors in which the nucleic acidsequences described herein are cloned into the vector in reverseorientation, but operably linked to a regulatory sequence that permitstranscription of antisense RNA. Thus, an antisense transcript can beproduced to all, or to a portion, of the nucleic acid molecule sequencesdescribed herein, including both coding and non-coding regions.Expression of this antisense RNA is subject to each of the parametersdescribed above in relation to expression of the sense RNA (regulatorysequences, constitutive or inducible expression, tissue-specificexpression).

[0188] The invention also relates to recombinant host cells containingthe vectors described herein. Host cells therefore include prokaryoticcells, lower e-karyotic cells such as yeast, other eukaryotic cells suchas insect cells, and higher eukaryotic cells such as mammalian cells.

[0189] The recombinant host cells are prepared by introducing the vectorconstructs described herein into the cells by techniques readilyavailable to the person of ordinary skill in the art. These include, butare not limited to, calcium phosphate transfection,DEAE-dextran-mediated transfection, cationic lipid-mediatedtransfection, electroporation, transduction, infection, lipofection, andother techniques such as those found in Sambrook, et al. (MolecularCloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).

[0190] Host cells can contain more than one vector. Thus, differentnucleotide sequences can be introduced on different vectors of the samecell. Similarly, the nucleic acid molecules can be introduced eitheralone or with other nucleic acid molecules that are not related to thenucleic acid molecules such as those providing trans-acting factors forexpression vectors. When more than one vector is introduced into a cell,the vectors can be introduced independently, co-introduced or joined tothe nucleic acid molecule vector.

[0191] In the case of bacteriophage and viral vectors, these can beintroduced into cells as packaged or encapsulated virus by standardprocedures for infection and transduction. Viral vectors can bereplication-competent or replication-defective. In the case in whichviral replication is defective, replication will occur in host cellsproviding functions that complement the defects.

[0192] Vectors generally include selectable markers that enable theselection of the subpopulation of cells that contain the recombinantvector constructs. The marker can be contained in the same vector thatcontains the nucleic acid molecules described herein or may be on aseparate vector. Markers include tetracycline or ampicillin-resistancegenes for prokaryotic host cells and dihydrofolate reductase or neomycinresistance for eukaryotic host cells. However, any marker that providesselection for a phenotypic trait will be effective.

[0193] While the mature proteins can be produced in bacteria, yeast,mammalian cells, and other cells under the control of the appropriateregulatory sequences, cell-free transcription and translation systemscan also be used to produce these proteins using RNA derived from theDNA constructs described herein.

[0194] Where secretion of the peptide is desired, which is difficult toachieve with multi-transmembrane domain containing proteins such askinases, appropriate secretion signals are incorporated into the vector.The signal sequence can be endogenous to the peptides or heterologous tothese peptides.

[0195] Where the peptide is not secreted into the medium, which istypically the case with kinases, the protein can be isolated from thehost cell by standard disruption procedures, including freeze thaw,sonication, mechanical disruption, use of lysing agents and the like.The peptide can then be recovered and purified by well-knownpurification methods including ammonium sulfate precipitation, acidextraction, anion or cationic exchange chromatography, phosphocellulosechromatography, hydrophobic-interaction chromatography, affinitychromatography, hydroxylapatite chromatography, lectin chromatography,or high performance liquid chromatography.

[0196] It is also understood that depending upon the host cell inrecombinant production of the peptides described herein, the peptidescan have various glycosylation patterns, depending upon the cell, ormaybe non-glycosylated as when produced in bacteria. In addition, thepeptides may include an initial modified methionine in some cases as aresult of a host-mediated process.

[0197] Uses of vectors and host cells

[0198] The recombinant host cells expressing the peptides describedherein have a variety of uses. First, the cells are useful for producinga kinase protein or peptide that can be further purified to producedesired amounts of kinase protein or fragments. Thus, host cellscontaining expression vectors are useful for peptide production.

[0199] Host cells are also useful for conducting cell-based assaysinvolving the kinase protein or kinase protein fragments, such as thosedescribed above as well as other formats known in the art. Thus, arecombinant host cell expressing a native kinase protein is useful forassaying compounds that stimulate or inhibit kinase protein function.

[0200] Host cells are also useful for identifying kinase protein mutantsin which these functions are affected. If the mutants naturally occurand give rise to a pathology, host cells containing the mutations areuseful to assay compounds that have a desired effect on the mutantkinase protein (for example, stimulating or inhibiting function which)may not be indicated by their effect on the native kinase protein.

[0201] Genetically engineered host cells can be further used to producenon-human transgenic animals. A transgenic animal is preferably amammal, for example a rodent, such as a rat or mouse, in which one ormore of the cells of the animal include a transgene. A transgene isexogenous DNA which is integrated into the genome of a cell from which atransgenic animal develops and which remains in the genome of the matureanimal in one or more cell types or tissues of the transgenic animal.These animals are useful for studying the function of a kinase proteinand identifying and evaluating modulators of kinase protein activity.Other examples of transgenic animals include non-human primates, sheep,dogs, cows, goats, chickens, and amphibians.

[0202] A transgenic animal can be produced by introducing nucleic acidinto the male pronuclei of a fertilized oocyte, e.g., by microinjection,retroviral infection, and allowing the oocyte to develop in apseudopregnant female foster animal. Any of the kinase proteinnucleotide sequences can be introduced as a transgene into the genome ofa non-human animal, such as a mouse.

[0203] Any of the regulatory or other sequences useful in expressionvectors can form part of the transgenic sequence. This includes intronicsequences and polyadenylation signals, if not already included. Atissue-specific regulatory sequence(s) can be operably linked to thetransgene to direct expression of the kinase protein to particularcells.

[0204] Methods for generating transgenic animals via embryo manipulationand microinjection, particularly animals such as mice, have becomeconventional in the art and are described, for example, in U.S. Pat.Nos. 4,736,866 and 4,870,009, both by Leder et al., U.S. Pat. No.4,873,191 by Wagner et al. and in Hogan, B., Manipulating the MouseEmbryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.,1986). Similar methods are used for production of other transgenicanimals. A transgenic founder animal can be identified based upon thepresence of the transgene in its genome and/or expression of transgenicmRNA in tissues or cells of the animals. A transgenic founder animal canthen be used to breed additional animals carrying the transgene.Moreover, transgenic animals carrying a transgene can further be bred toother transgenic animals carrying other transgenes. A transgenic animalalso includes animals in which the entire animal or tissues in theanimal have been produced using the homologously recombinant host Cellsdescribed herein.

[0205] In another embodiment, transgenic non-human animals can beproduced which contain selected systems that allow for regulatedexpression of the transgene. One example of such a system is thecre/loxP recombinase system of bacteriophage P1. For a description ofthe cre/loxP recombinase system, see, e.g., Lakso et al. PNAS89:6232-6236 (1992). Another example of a recombinase system is the FLPrecombinase system of S. cerevisiae (O'Gorman et al. Science251:1351-1355 (1991). If a cre/loxP recombinase system is used toregulate expression of the transgene, animals containing transgenesencoding both the Cre recombinase and a selected protein is required.Such animals can be provided through the construction of “double”transgenic animals, e.g., by mating two transgenic animals, onecontaining a transgene encoding a selected protein and the othercontaining a transgene encoding a recombinase.

[0206] Clones of the non-human transgenic animals described herein canalso be produced according to the methods described in Wilmut, I. et al.Nature 385:810-813 (1997) and PCT International Publication Nos. WO97/07668 and WO 97/07669. In brief, a cell, e.g., a somatic cell, fromthe transgenic animal can be isolated and induced to exit the growthcycle and enter G_(o) phase. The quiescent cell can then be fused, e.g.,through the use of electrical pulses, to an enucleated oocyte from ananimal of the same species from which the quiescent cell is isolated.The reconstructed oocyte is then cultured such that it develops tomorula or blastocyst and then transferred to pseudopregnant femalefoster animal. The offspring born of this female foster animal will be aclone of the animal from which the cell, e.g., the somatic cell, isisolated.

[0207] Transgenic animals containing recombinant cells that express thepeptides described herein are useful to conduct the assays describedherein in an in vivo context. Accordingly, the various physiologicalfactors that are present in vivo and that could effect substratebinding, kinase protein activation, and signal transduction, may not beevident from in vitro cell-free or cell-based assays. Accordingly, it isuseful to provide non-human transgenic animals to assay in vivo kinaseprotein function, including substrate interaction, the effect ofspecific mutant kinase proteins on kinase protein function and substrateinteraction, and the effect of chimeric kinase proteins. It is alsopossible to assess the effect of null mutations, that is, mutations thatsubstantially or completely eliminate one or more kinase proteinfunctions.

[0208] All publications and patents mentioned in the above specificationare herein incorporated by reference. Various modifications andvariations of the described method and system of the invention will beapparent to those skilled in the art without departing from the scopeand spirit of the invention. Although the invention has been describedin connection with specific preferred embodiments, it should beunderstood that the invention as claimed should not be unduly limited tosuch specific embodiments. Indeed, various modifications of theabove-described modes for carrying out the invention which are obviousto those skilled in the field of molecular biology or related fields areintended to be within the scope of the following claims.

1 34 1 5207 DNA Homo sapiens 1 cagcacgagg aactccttct gatcacctggccagctgagg tcagagtggg agaggcagtg 60 gttccattga aggagtactc ctaactgtcagaagcctggg cggtcaggat ggggtgctgt 120 cgcttgggct gcggggggtg ttcagttgcccacagtgtat ctcagggtct caccaaccat 180 ccaagcatgg taggctgtgg ctggcacccagggttgtgtg gctggggagg tggtctccac 240 agttccctcc ctgccctccc agggcccccatccatgcagg taaccatcga ggatgtgcag 300 gcacagacag gcggaacggc ccaattcgaggctatcattg agggcgaccc acagccctcg 360 gtgacctggt acaaggacag cgtccagctggtggacagca cccggcttag ccagcagcaa 420 gaaggcacca catactccct ggtgctgaggcatgtggcct cgaaggatgc cggcgtttac 480 acctgcctgg cccaaaacac tggtggccaggtgctctgca aggcagagct gctggtgctt 540 gggggggaca atgagccgga ctcagagaagcaaagccacc ggaggaagct gcactccttc 600 tatgaggtca aggaggagat tggaaggggcgtgtttggct tcgtaaaaag agtgcagcac 660 aaaggaaaca agatcttgtg cgctgccaagttcatccccc tacggagcag aactcgggcc 720 caggcataca gggagcgaga catcctggccgcgctgagcc acccgctggt cacggggctg 780 ctggaccagt ttgagacccg caagaccctcatcctcatcc tggagctgtg ctcatccgag 840 gagctgctgg accgcctgta caggaagggcgtggtgacgg aggccgaggt caaggtctac 900 atccagcagc tggtggaggg gctgcactacctgcacagcc atggcgttct ccacctggac 960 ataaagccct ctaacatcct gatggtgcatcctgcccggg aagacattaa aatctgcgac 1020 tttggctttg cccagaacat caccccagcagagctgcagt tcagccagta cggctcccct 1080 gagttcgtct cccccgagat catccagcagaaccctgtga gcgaagcctc cgacatttgg 1140 gccatgggtg tcatctccta cctcagcctgacctgctcat ccccatttgc cggcgagagt 1200 gaccgtgcca ccctcctgaa cgtcctggaggggcgcgtgt catggagcag ccccatggct 1260 gcccacctca gcgaagacgc caaagacttcatcaaggcta cgctgcagag agcccctcag 1320 gcccggccta gtgcggccca gtgcctctcccacccctggt tcctgaaatc catgcctgcg 1380 gaggaggccc acttcatcaa caccaagcagctcaagttcc tcctggcccg aagtcgctgg 1440 cagcgttccc tgatgagcta caagtccatcctggtgatgc gctccatccc tgagctgctg 1500 cggggcccac ccgacagccc ctccctcggcgtagcccggc acctctgcag ggacactggt 1560 ggctcctcca gttcctcctc ctcctctgacaacgagctcg ccccatttgc ccgggctaag 1620 tcactgccac cctccccggt gacacactcaccactgctgc acccccgggg cttcctgcgg 1680 ccctcggcca gcctgcctga ggaagccgaggccagtgagc gctccaccga ggccccagct 1740 ccgcctgcat ctcccgaggg tgccgggccaccggccgccc agggctgcgt gccccggcac 1800 agcgtcatcc gcagcctgtt ctaccaccaggcgggtgaga gccctgagca cggggccctg 1860 gccccgggga gcaggcggca cccggcccggcggcggcacc tgctgaaggg cggctacatt 1920 gcgggggcgc tgccaggcct gcgcgagccactgatggagc accgcgtgct ggaggaggag 1980 gccgccaggg aggagcaggc caccctcctggccaaagccc cctcattcga gactgccctc 2040 cggctgcctg cctctggcac ccacttggcccctggccaca gccactccct ggaacatgac 2100 tctccgagca ccccccgccc ctcctcggaggcctgcggtg aggcacagcg actgccttca 2160 gccccctccg ggggggcccc tatcagggacatggggcacc ctcagggctc caagcagctt 2220 ccatccactg gtggccaccc aggcactgctcagccagaga ggccatcccc ggacagccct 2280 tgggggcagc cagccccttt ctgccaccccaagcagggtt ctgcccccca ggagggctgc 2340 agcccccacc cagcagttgc cccatgccctcctggctcct tccctccagg atcttgcaaa 2400 gaggccccct tagtaccctc aagccccttcttgggacagc cccaggcacc ccctgcccct 2460 gccaaagcaa gccccccatt ggactctaagatggggcctg gagacatctc tcttcctggg 2520 aggccaaaac ccggcccctg cagttccccagggtcagcct cccaggcgag ctcttcccaa 2580 gtgagctccc tcagggtggg ctcctcccaggtgggcacag agcctggccc ctccctggat 2640 gcggagggct ggacccagga ggctgaggatctgtccgact ccacacccac cttgcagcgg 2700 cctcaggaac aggtgaccat gcgcaagttctccctgggtg gtcgcggggg ctacgcaggc 2760 gtggctggct atggcacctt tgcctttggtggagatgcag ggggcatgct ggggcagggg 2820 cccatgtggg ccaggatagc ctgggctgtgtcccagtcgg aggaggagga gcaggaggag 2880 gccagggctg agtcccagtc ggaggagcagcaggaggcca gggctgagag cccactgccc 2940 caggtcagtg caaggcctgt gcctgaggtcggcagggctc ccaccaggag ctctccagag 3000 cccaccccat gggaggacat cgggcaggtctccctggtgc agatccggga cctgtcaggt 3060 gatgcggagg cggccgacac aatatccctggacatttccg aggtggaccc cgcctacctc 3120 aacctctcag acctgtacga tatcaagtacctcccattcg agtttatgat cttcaggaaa 3180 gtccccaagt ccgctcagcc agagccgccctcccccatgg ctgaggagga gctggccgag 3240 ttcccggagc ccacgtggcc ctggccaggtgaactgggcc cccacgcagg cctggagatc 3300 acagaggagt cagaggatgt ggacgcgctgctggcagagg ctgccgtggg caggaagcgc 3360 aagtggtcct cgccgtcacg cagcctcttccacttccctg ggaggcacct gccgctggat 3420 gagcctgcag agctggggct gcgtgagagagtgaaggcct ccgtggagca catctcccgg 3480 atcctgaagg gcaggccgga aggtctggagaaggaggggc cccccaggaa gaagccaggc 3540 cttgcttcct tccggctctc aggtctgaagagctgggacc gagcgccgac attcctaagg 3600 gagctctcag atgagactgt ggtcctgggccagtcagtga cactggcctg ccaggtgtca 3660 gcccagccag ctgcccaggc cacctggagcaaagacggag cccccctgga gagcagcagc 3720 cgtgtcctca tctctgccac cctcaagaacttccagcttc tgaccatcct ggtggtggtg 3780 gctgaggacc tgggtgtgta cacctgcagcgtgagcaatg cgctggggac agtgaccacc 3840 acgggcgtcc tccggaaggc agagcgcccctcatcttcgc catgcccgga tatcggggag 3900 gtgtacgcgg atggggtgct gctggtctggaagcccgtgg aatcctacgg ccctgtgacc 3960 tacattgtgc agtgcagcct agaaggcggcagctggacca cactggcctc cgacatcttt 4020 gactgctgct acctgaccag caagctctcccggggtggca cctacacctt ccgcacggca 4080 tgtgtcagca aggcaggaat gggtccctacagcagcccct cggagcaagt cctcctggga 4140 gggcccagcc acctggcctc tgaggaggagagccaggggc ggtcagccca acccctgccc 4200 agcacaaaga ccttcgcatt ccagacacagatccagaggg gccgcttcag cgtggtgcgg 4260 caatgctggg agaaggccag cgggcgggcgctggccgcca agatcatccc ctaccacccc 4320 aaggacaaga cagcagtgct gcgcgaatacgaggccctca agggcctgcg ccacccgcac 4380 ctggcccagc tgcacgcagc ctacctcagcccccggcacc tggtgctcat cttggagctg 4440 tgctctgggc ccgagctgct cccctgcctggccgagaggg cctcctactc agaatctgag 4500 gtgaaggact acctgtggca gatgttgagtgccacccagt acctgcacaa ccagcacatc 4560 ctgcacctgg acctgaggtc cgagaacatgatcatcaccg aatacaacct gctcaaggtc 4620 gtggacctgg gcaatgcaca gagcctcagccaggagaagg tgctgccctc agacaagttc 4680 aaggactacc tagagaccat ggctccagagctcctggagg gccagggggc tgttccacag 4740 acagacatct gggccatcgg tgtgacagccttcatcatgc tgagcgccga gtacccggtg 4800 agcagcgagg gtgcacgcga cctgcagagaggactgcgca aggggctggt ccggctgagc 4860 cgctgctacg cggggctgtc cgggggcgccgtggccttcc tgcgcagcac tctgtgcgcc 4920 cagccctggg gccggccctg cgcgtccagctgcctgcagt gcccgtggct aacagaggag 4980 ggcccggcct gttcgcggcc cgcgcccgtgaccttcccta ccgcgcggct gcgcgtcttc 5040 gtgcgcaatc gcgagaagag acgcgcgctgctgtacaaga ggcacaacct ggcccaggtg 5100 cgctgagggt cgccccggcc acacccttggtctccccgct gggggtcgct gcagacgcgc 5160 caataaaaac gcacagccgg gcgagaaaaaaaaaaaaaaa aaaaaaa 5207 2 1665 PRT Homo sapiens 2 Met Gly Cys Cys ArgLeu Gly Cys Gly Gly Cys Ser Val Ala His Ser 1 5 10 15 Val Ser Gln GlyLeu Thr Asn His Pro Ser Met Val Gly Cys Gly Trp 20 25 30 His Pro Gly LeuCys Gly Trp Gly Gly Gly Leu His Ser Ser Leu Pro 35 40 45 Ala Leu Pro GlyPro Pro Ser Met Gln Val Thr Ile Glu Asp Val Gln 50 55 60 Ala Gln Thr GlyGly Thr Ala Gln Phe Glu Ala Ile Ile Glu Gly Asp 65 70 75 80 Pro Gln ProSer Val Thr Trp Tyr Lys Asp Ser Val Gln Leu Val Asp 85 90 95 Ser Thr ArgLeu Ser Gln Gln Gln Glu Gly Thr Thr Tyr Ser Leu Val 100 105 110 Leu ArgHis Val Ala Ser Lys Asp Ala Gly Val Tyr Thr Cys Leu Ala 115 120 125 GlnAsn Thr Gly Gly Gln Val Leu Cys Lys Ala Glu Leu Leu Val Leu 130 135 140Gly Gly Asp Asn Glu Pro Asp Ser Glu Lys Gln Ser His Arg Arg Lys 145 150155 160 Leu His Ser Phe Tyr Glu Val Lys Glu Glu Ile Gly Arg Gly Val Phe165 170 175 Gly Phe Val Lys Arg Val Gln His Lys Gly Asn Lys Ile Leu CysAla 180 185 190 Ala Lys Phe Ile Pro Leu Arg Ser Arg Thr Arg Ala Gln AlaTyr Arg 195 200 205 Glu Arg Asp Ile Leu Ala Ala Leu Ser His Pro Leu ValThr Gly Leu 210 215 220 Leu Asp Gln Phe Glu Thr Arg Lys Thr Leu Ile LeuIle Leu Glu Leu 225 230 235 240 Cys Ser Ser Glu Glu Leu Leu Asp Arg LeuTyr Arg Lys Gly Val Val 245 250 255 Thr Glu Ala Glu Val Lys Val Tyr IleGln Gln Leu Val Glu Gly Leu 260 265 270 His Tyr Leu His Ser His Gly ValLeu His Leu Asp Ile Lys Pro Ser 275 280 285 Asn Ile Leu Met Val His ProAla Arg Glu Asp Ile Lys Ile Cys Asp 290 295 300 Phe Gly Phe Ala Gln AsnIle Thr Pro Ala Glu Leu Gln Phe Ser Gln 305 310 315 320 Tyr Gly Ser ProGlu Phe Val Ser Pro Glu Ile Ile Gln Gln Asn Pro 325 330 335 Val Ser GluAla Ser Asp Ile Trp Ala Met Gly Val Ile Ser Tyr Leu 340 345 350 Ser LeuThr Cys Ser Ser Pro Phe Ala Gly Glu Ser Asp Arg Ala Thr 355 360 365 LeuLeu Asn Val Leu Glu Gly Arg Val Ser Trp Ser Ser Pro Met Ala 370 375 380Ala His Leu Ser Glu Asp Ala Lys Asp Phe Ile Lys Ala Thr Leu Gln 385 390395 400 Arg Ala Pro Gln Ala Arg Pro Ser Ala Ala Gln Cys Leu Ser His Pro405 410 415 Trp Phe Leu Lys Ser Met Pro Ala Glu Glu Ala His Phe Ile AsnThr 420 425 430 Lys Gln Leu Lys Phe Leu Leu Ala Arg Ser Arg Trp Gln ArgSer Leu 435 440 445 Met Ser Tyr Lys Ser Ile Leu Val Met Arg Ser Ile ProGlu Leu Leu 450 455 460 Arg Gly Pro Pro Asp Ser Pro Ser Leu Gly Val AlaArg His Leu Cys 465 470 475 480 Arg Asp Thr Gly Gly Ser Ser Ser Ser SerSer Ser Ser Asp Asn Glu 485 490 495 Leu Ala Pro Phe Ala Arg Ala Lys SerLeu Pro Pro Ser Pro Val Thr 500 505 510 His Ser Pro Leu Leu His Pro ArgGly Phe Leu Arg Pro Ser Ala Ser 515 520 525 Leu Pro Glu Glu Ala Glu AlaSer Glu Arg Ser Thr Glu Ala Pro Ala 530 535 540 Pro Pro Ala Ser Pro GluGly Ala Gly Pro Pro Ala Ala Gln Gly Cys 545 550 555 560 Val Pro Arg HisSer Val Ile Arg Ser Leu Phe Tyr His Gln Ala Gly 565 570 575 Glu Ser ProGlu His Gly Ala Leu Ala Pro Gly Ser Arg Arg His Pro 580 585 590 Ala ArgArg Arg His Leu Leu Lys Gly Gly Tyr Ile Ala Gly Ala Leu 595 600 605 ProGly Leu Arg Glu Pro Leu Met Glu His Arg Val Leu Glu Glu Glu 610 615 620Ala Ala Arg Glu Glu Gln Ala Thr Leu Leu Ala Lys Ala Pro Ser Phe 625 630635 640 Glu Thr Ala Leu Arg Leu Pro Ala Ser Gly Thr His Leu Ala Pro Gly645 650 655 His Ser His Ser Leu Glu His Asp Ser Pro Ser Thr Pro Arg ProSer 660 665 670 Ser Glu Ala Cys Gly Glu Ala Gln Arg Leu Pro Ser Ala ProSer Gly 675 680 685 Gly Ala Pro Ile Arg Asp Met Gly His Pro Gln Gly SerLys Gln Leu 690 695 700 Pro Ser Thr Gly Gly His Pro Gly Thr Ala Gln ProGlu Arg Pro Ser 705 710 715 720 Pro Asp Ser Pro Trp Gly Gln Pro Ala ProPhe Cys His Pro Lys Gln 725 730 735 Gly Ser Ala Pro Gln Glu Gly Cys SerPro His Pro Ala Val Ala Pro 740 745 750 Cys Pro Pro Gly Ser Phe Pro ProGly Ser Cys Lys Glu Ala Pro Leu 755 760 765 Val Pro Ser Ser Pro Phe LeuGly Gln Pro Gln Ala Pro Pro Ala Pro 770 775 780 Ala Lys Ala Ser Pro ProLeu Asp Ser Lys Met Gly Pro Gly Asp Ile 785 790 795 800 Ser Leu Pro GlyArg Pro Lys Pro Gly Pro Cys Ser Ser Pro Gly Ser 805 810 815 Ala Ser GlnAla Ser Ser Ser Gln Val Ser Ser Leu Arg Val Gly Ser 820 825 830 Ser GlnVal Gly Thr Glu Pro Gly Pro Ser Leu Asp Ala Glu Gly Trp 835 840 845 ThrGln Glu Ala Glu Asp Leu Ser Asp Ser Thr Pro Thr Leu Gln Arg 850 855 860Pro Gln Glu Gln Val Thr Met Arg Lys Phe Ser Leu Gly Gly Arg Gly 865 870875 880 Gly Tyr Ala Gly Val Ala Gly Tyr Gly Thr Phe Ala Phe Gly Gly Asp885 890 895 Ala Gly Gly Met Leu Gly Gln Gly Pro Met Trp Ala Arg Ile AlaTrp 900 905 910 Ala Val Ser Gln Ser Glu Glu Glu Glu Gln Glu Glu Ala ArgAla Glu 915 920 925 Ser Gln Ser Glu Glu Gln Gln Glu Ala Arg Ala Glu SerPro Leu Pro 930 935 940 Gln Val Ser Ala Arg Pro Val Pro Glu Val Gly ArgAla Pro Thr Arg 945 950 955 960 Ser Ser Pro Glu Pro Thr Pro Trp Glu AspIle Gly Gln Val Ser Leu 965 970 975 Val Gln Ile Arg Asp Leu Ser Gly AspAla Glu Ala Ala Asp Thr Ile 980 985 990 Ser Leu Asp Ile Ser Glu Val AspPro Ala Tyr Leu Asn Leu Ser Asp 995 1000 1005 Leu Tyr Asp Ile Lys TyrLeu Pro Phe Glu Phe Met Ile Phe Arg Lys 1010 1015 1020 Val Pro Lys SerAla Gln Pro Glu Pro Pro Ser Pro Met Ala Glu Glu 1025 1030 1035 1040 GluLeu Ala Glu Phe Pro Glu Pro Thr Trp Pro Trp Pro Gly Glu Leu 1045 10501055 Gly Pro His Ala Gly Leu Glu Ile Thr Glu Glu Ser Glu Asp Val Asp1060 1065 1070 Ala Leu Leu Ala Glu Ala Ala Val Gly Arg Lys Arg Lys TrpSer Ser 1075 1080 1085 Pro Ser Arg Ser Leu Phe His Phe Pro Gly Arg HisLeu Pro Leu Asp 1090 1095 1100 Glu Pro Ala Glu Leu Gly Leu Arg Glu ArgVal Lys Ala Ser Val Glu 1105 1110 1115 1120 His Ile Ser Arg Ile Leu LysGly Arg Pro Glu Gly Leu Glu Lys Glu 1125 1130 1135 Gly Pro Pro Arg LysLys Pro Gly Leu Ala Ser Phe Arg Leu Ser Gly 1140 1145 1150 Leu Lys SerTrp Asp Arg Ala Pro Thr Phe Leu Arg Glu Leu Ser Asp 1155 1160 1165 GluThr Val Val Leu Gly Gln Ser Val Thr Leu Ala Cys Gln Val Ser 1170 11751180 Ala Gln Pro Ala Ala Gln Ala Thr Trp Ser Lys Asp Gly Ala Pro Leu1185 1190 1195 1200 Glu Ser Ser Ser Arg Val Leu Ile Ser Ala Thr Leu LysAsn Phe Gln 1205 1210 1215 Leu Leu Thr Ile Leu Val Val Val Ala Glu AspLeu Gly Val Tyr Thr 1220 1225 1230 Cys Ser Val Ser Asn Ala Leu Gly ThrVal Thr Thr Thr Gly Val Leu 1235 1240 1245 Arg Lys Ala Glu Arg Pro SerSer Ser Pro Cys Pro Asp Ile Gly Glu 1250 1255 1260 Val Tyr Ala Asp GlyVal Leu Leu Val Trp Lys Pro Val Glu Ser Tyr 1265 1270 1275 1280 Gly ProVal Thr Tyr Ile Val Gln Cys Ser Leu Glu Gly Gly Ser Trp 1285 1290 1295Thr Thr Leu Ala Ser Asp Ile Phe Asp Cys Cys Tyr Leu Thr Ser Lys 13001305 1310 Leu Ser Arg Gly Gly Thr Tyr Thr Phe Arg Thr Ala Cys Val SerLys 1315 1320 1325 Ala Gly Met Gly Pro Tyr Ser Ser Pro Ser Glu Gln ValLeu Leu Gly 1330 1335 1340 Gly Pro Ser His Leu Ala Ser Glu Glu Glu SerGln Gly Arg Ser Ala 1345 1350 1355 1360 Gln Pro Leu Pro Ser Thr Lys ThrPhe Ala Phe Gln Thr Gln Ile Gln 1365 1370 1375 Arg Gly Arg Phe Ser ValVal Arg Gln Cys Trp Glu Lys Ala Ser Gly 1380 1385 1390 Arg Ala Leu AlaAla Lys Ile Ile Pro Tyr His Pro Lys Asp Lys Thr 1395 1400 1405 Ala ValLeu Arg Glu Tyr Glu Ala Leu Lys Gly Leu Arg His Pro His 1410 1415 1420Leu Ala Gln Leu His Ala Ala Tyr Leu Ser Pro Arg His Leu Val Leu 14251430 1435 1440 Ile Leu Glu Leu Cys Ser Gly Pro Glu Leu Leu Pro Cys LeuAla Glu 1445 1450 1455 Arg Ala Ser Tyr Ser Glu Ser Glu Val Lys Asp TyrLeu Trp Gln Met 1460 1465 1470 Leu Ser Ala Thr Gln Tyr Leu His Asn GlnHis Ile Leu His Leu Asp 1475 1480 1485 Leu Arg Ser Glu Asn Met Ile IleThr Glu Tyr Asn Leu Leu Lys Val 1490 1495 1500 Val Asp Leu Gly Asn AlaGln Ser Leu Ser Gln Glu Lys Val Leu Pro 1505 1510 1515 1520 Ser Asp LysPhe Lys Asp Tyr Leu Glu Thr Met Ala Pro Glu Leu Leu 1525 1530 1535 GluGly Gln Gly Ala Val Pro Gln Thr Asp Ile Trp Ala Ile Gly Val 1540 15451550 Thr Ala Phe Ile Met Leu Ser Ala Glu Tyr Pro Val Ser Ser Glu Gly1555 1560 1565 Ala Arg Asp Leu Gln Arg Gly Leu Arg Lys Gly Leu Val ArgLeu Ser 1570 1575 1580 Arg Cys Tyr Ala Gly Leu Ser Gly Gly Ala Val AlaPhe Leu Arg Ser 1585 1590 1595 1600 Thr Leu Cys Ala Gln Pro Trp Gly ArgPro Cys Ala Ser Ser Cys Leu 1605 1610 1615 Gln Cys Pro Trp Leu Thr GluGlu Gly Pro Ala Cys Ser Arg Pro Ala 1620 1625 1630 Pro Val Thr Phe ProThr Ala Arg Leu Arg Val Phe Val Arg Asn Arg 1635 1640 1645 Glu Lys ArgArg Ala Leu Leu Tyr Lys Arg His Asn Leu Ala Gln Val 1650 1655 1660 Arg1665 3 5207 DNA Homo sapiens 3 cagcacgagg aactccttct gatcacctggccagctgagg tcagagtggg agaggcagtg 60 gttccattga aggagtactc ctaactgtcagaagcctggg cggtcaggat ggggtgctgt 120 cgcttgggct gcggggggtg ttcagttgcccacagtgtat ctcagggtct caccaaccat 180 ccaagcatgg taggctgtgg ctggcacccagggttgtgtg gctggggagg tggtctccac 240 agttccctcc ctgccctccc agggcccccatccatgcagg taaccatcga ggatgtgcag 300 gcacagacag gcggaacggc ccaattcgaggctatcattg agggcgaccc acagccctcg 360 gtgacctggt acaaggacag cgtccagctggtggacagca cccggcttag ccagcagcaa 420 gaaggcacca catactccct ggtgctgaggcatgtggcct cgaaggatgc cggcgtttac 480 acctgcctgg cccaaaacac tggtggccaggtgctctgca aggcagagct gctggtgctt 540 gggggggaca atgagccgga ctcagagaagcaaagccacc ggaggaagct gcactccttc 600 tatgaggtca aggaggagat tggaaggggcgtgtttggct tcgtaaaaag agtgcagcac 660 aaaggaaaca agatcttgtg cgctgccaagttcatccccc tacggagcag aactcgggcc 720 caggcataca gggagcgaga catcctggccgcgctgagcc acccgctggt cacggggctg 780 ctggaccagt ttgagacccg caagaccctcatcctcatcc tggagctgtg ctcatccgag 840 gagctgctgg accgcctgta caggaagggcgtggtgacgg aggccgaggt caaggtctac 900 atccagcagc tggtggaggg gctgcactacctgcacagcc atggcgttct ccacctggac 960 ataaagccct ctaacatcct gatggtgcatcctgcccggg aagacattaa aatctgcgac 1020 tttggctttg cccagaacat caccccagcagagctgcagt tcagccagta cggctcccct 1080 gagttcgtct cccccgagat catccagcagaaccctgtga gcgaagcctc cgacatttgg 1140 gccatgggtg tcatctccta cctcagcctgacctgctcat ccccatttgc cggcgagagt 1200 gaccgtgcca ccctcctgaa cgtcctggaggggcgcgtgt catggagcag ccccatggct 1260 gcccacctca gcgaagacgc caaagacttcatcaaggcta cgctgcagag agcccctcag 1320 gcccggccta gtgcggccca gtgcctctcccacccctggt tcctgaaatc catgcctgcg 1380 gaggaggccc acttcatcaa caccaagcagctcaagttcc tcctggcccg aagtcgctgg 1440 cagcgttccc tgatgagcta caagtccatcctggtgatgc gctccatccc tgagctgctg 1500 cggggcccac ccgacagccc ctccctcggcgtagcccggc acctctgcag ggacactggt 1560 ggctcctcca gttcctcctc ctcctctgacaacgagctcg ccccatttgc ccgggctaag 1620 tcactgccac cctccccggt gacacactcaccactgctgc acccccgggg cttcctgcgg 1680 ccctcggcca gcctgcctga ggaagccgaggccagtgagc gctccaccga ggccccagct 1740 ccgcctgcat ctcccgaggg tgccgggccaccggccgccc agggctgcgt gccccggcac 1800 agcgtcatcc gcagcctgtt ctaccaccaggcgggtgaga gccctgagca cggggccctg 1860 gccccgggga gcaggcggca cccggcccggcggcggcacc tgctgaaggg cggctacatt 1920 gcgggggcgc tgccaggcct gcgcgagccactgatggagc accgcgtgct ggaggaggag 1980 gccgccaggg aggagcaggc caccctcctggccaaagccc cctcattcga gactgccctc 2040 cggctgcctg cctctggcac ccacttggcccctggccaca gccactccct ggaacatgac 2100 tctccgagca ccccccgccc ctcctcggaggcctgcggtg aggcacagcg actgccttca 2160 gccccctccg ggggggcccc tatcagggacatggggcacc ctcagggctc caagcagctt 2220 ccatccactg gtggccaccc aggcactgctcagccagaga ggccatcccc ggacagccct 2280 tgggggcagc cagccccttt ctgccaccccaagcagggtt ctgcccccca ggagggctgc 2340 agcccccacc cagcagttgc cccatgccctcctggctcct tccctccagg atcttgcaaa 2400 gaggccccct tagtaccctc aagccccttcttgggacagc cccaggcacc ccctgcccct 2460 gccaaagcaa gccccccatt ggactctaagatggggcctg gagacatctc tcttcctggg 2520 aggccaaaac ccggcccctg cagttccccagggtcagcct cccaggcgag ctcttcccaa 2580 gtgagctccc tcagggtggg ctcctcccaggtgggcacag agcctggccc ctccctggat 2640 gcggagggct ggacccagga ggctgaggatctgtccgact ccacacccac cttgcagcgg 2700 cctcaggaac aggtgaccat gcgcaagttctccctgggtg gtcgcggggg ctacgcaggc 2760 gtggctggct atggcacctt tgcctttggtggagatgcag ggggcatgct ggggcagggg 2820 cccatgtggg ccaggatagc ctgggctgtgtcccagtcgg aggaggagga gcaggaggag 2880 gccagggctg agtcccagtc ggaggagcagcaggaggcca gggctgagag cccactgccc 2940 caggtcagtg caaggcctgt gcctgaggtcggcagggctc ccaccaggag ctctccagag 3000 cccaccccat gggaggacat cgggcaggtctccctggtgc agatccggga cctgtcaggt 3060 gatgcggagg cggccgacac aatatccctggacatttccg aggtggaccc cgcctacctc 3120 aacctctcag acctgtacga tatcaagtacctcccattcg agtttatgat cttcaggaaa 3180 gtccccaagt ccgctcagcc agagccgccctcccccatgg ctgaggagga gctggccgag 3240 ttcccggagc ccacgtggcc ctggccaggtgaactgggcc cccacgcagg cctggagatc 3300 acagaggagt cagaggatgt ggacgcgctgctggcagagg ctgccgtggg caggaagcgc 3360 aagtggtcct cgccgtcacg cagcctcttccacttccctg ggaggcacct gccgctggat 3420 gagcctgcag agctggggct gcgtgagagagtgaaggcct ccgtggagca catctcccgg 3480 atcctgaagg gcaggccgga aggtctggagaaggaggggc cccccaggaa gaagccaggc 3540 cttgcttcct tccggctctc aggtctgaagagctgggacc gagcgccgac attcctaagg 3600 gagctctcag atgagactgt ggtcctgggccagtcagtga cactggcctg ccaggtgtca 3660 gcccagccag ctgcccaggc cacctggagcaaagacggag cccccctgga gagcagcagc 3720 cgtgtcctca tctctgccac cctcaagaacttccagcttc tgaccatcct ggtggtggtg 3780 gctgaggacc tgggtgtgta cacctgcagcgtgagcaatg cgctggggac agtgaccacc 3840 acgggcgtcc tccggaaggc agagcgcccctcatcttcgc catgcccgga tatcggggag 3900 gtgtacgcgg atggggtgct gctggtctggaagcccgtgg aatcctacgg ccctgtgacc 3960 tacattgtgc agtgcagcct agaaggcggcagctggacca cactggcctc cgacatcttt 4020 gactgctgct acctgaccag caagctctcccggggtggca cctacacctt ccgcacggca 4080 tgtgtcagca aggcaggaat gggtccctacagcagcccct cggagcaagt cctcctggga 4140 gggcccagcc acctggcctc tgaggaggagagccaggggc ggtcagccca acccctgccc 4200 agcacaaaga ccttcgcatt ccagacacagatccagaggg gccgcttcag cgtggtgcgg 4260 caatgctggg agaaggccag cgggcgggcgctggccgcca agatcatccc ctaccacccc 4320 aaggacaaga cagcagtgct gcgcgaatacgaggccctca agggcctgcg ccacccgcac 4380 ctggcccagc tgcacgcagc ctacctcagcccccggcacc tggtgctcat cttggagctg 4440 tgctctgggc ccgagctgct cccctgcctggccgagaggg cctcctactc agaatctgag 4500 gtgaaggact acctgtggca gatgttgagtgccacccagt acctgcacaa ccagcacatc 4560 ctgcacctgg acctgaggtc cgagaacatgatcatcaccg aatacaacct gctcaaggtc 4620 gtggacctgg gcaatgcaca gagcctcagccaggagaagg tgctgccctc agacaagttc 4680 aaggactacc tagagaccat ggctccagagctcctggagg gccagggggc tgttccacag 4740 acagacatct gggccatcgg tgtgacagccttcatcatgc tgagcgccga gtacccggtg 4800 agcagcgagg gtgcacgcga cctgcagagaggactgcgca aggggctggt ccggctgagc 4860 cgctgctacg cggggctgtc cgggggcgccgtggccttcc tgcgcagcac tctgtgcgcc 4920 cagccctggg gccggccctg cgcgtccagctgcctgcagt gcccgtggct aacagaggag 4980 ggcccggcct gttcgcggcc cgcgcccgtgaccttcccta ccgcgcggct gcgcgtcttc 5040 gtgcgcaatc gcgagaagag acgcgcgctgctgtacaaga ggcacaacct ggcccaggtg 5100 cgctgagggt cgccccggcc acacccttggtctccccgct gggggtcgct gcagacgcgc 5160 caataaaaac gcacagccgg gcgagaaaaaaaaaaaaaaa aaaaaaa 5207 4 846 PRT Homo sapiens 4 Pro Arg Phe Glu Ser IleMet Glu Asp Val Glu Val Gly Ala Gly Glu 1 5 10 15 Thr Ala Arg Phe AlaVal Val Val Glu Gly Lys Pro Leu Pro Asp Ile 20 25 30 Met Trp Tyr Lys AspGlu Val Leu Leu Thr Glu Ser Ser His Val Ser 35 40 45 Phe Val Tyr Glu GluAsn Glu Cys Ser Leu Val Val Leu Ser Thr Gly 50 55 60 Ala Gln Asp Gly GlyVal Tyr Thr Cys Thr Ala Gln Asn Leu Ala Gly 65 70 75 80 Glu Val Ser CysLys Ala Glu Leu Ala Val His Ser Ala Gln Thr Ala 85 90 95 Met Glu Val GluGly Val Gly Glu Asp Glu Asp His Arg Gly Arg Arg 100 105 110 Leu Ser AspPhe Tyr Asp Ile His Gln Glu Ile Gly Arg Gly Ala Phe 115 120 125 Ser TyrLeu Arg Arg Ile Val Glu Arg Ser Ser Gly Leu Glu Phe Ala 130 135 140 AlaLys Phe Ile Pro Ser Gln Ala Lys Pro Lys Ala Ser Ala Arg Arg 145 150 155160 Glu Ala Arg Leu Leu Ala Arg Leu Gln His Asp Cys Val Leu Tyr Phe 165170 175 His Glu Ala Phe Glu Arg Arg Arg Gly Leu Val Ile Val Thr Glu Leu180 185 190 Cys Thr Glu Glu Leu Leu Glu Arg Ile Ala Arg Lys Pro Thr ValCys 195 200 205 Glu Ser Glu Ile Arg Ala Tyr Met Arg Gln Val Leu Glu GlyIle His 210 215 220 Tyr Leu His Gln Ser His Val Leu His Leu Asp Val LysPro Glu Asn 225 230 235 240 Leu Leu Val Trp Asp Gly Ala Ala Gly Glu GlnGln Val Arg Ile Cys 245 250 255 Asp Phe Gly Asn Ala Gln Glu Leu Thr ProGly Glu Pro Gln Tyr Cys 260 265 270 Gln Tyr Gly Thr Pro Glu Phe Val AlaPro Glu Ile Val Asn Gln Ser 275 280 285 Pro Val Ser Gly Val Thr Asp IleTrp Pro Val Gly Val Val Ala Phe 290 295 300 Leu Cys Leu Thr Gly Ile SerPro Phe Val Gly Glu Asn Asp Arg Thr 305 310 315 320 Thr Leu Met Asn IleArg Asn Tyr Asn Val Ala Phe Glu Glu Thr Thr 325 330 335 Phe Leu Ser LeuSer Arg Glu Ala Arg Gly Phe Leu Ile Lys Val Leu 340 345 350 Val Gln AspArg Leu Arg Pro Thr Ala Glu Glu Thr Leu Glu His Pro 355 360 365 Trp PheLys Thr Gln Ala Lys Gly Ala Glu Val Ser Thr Asp His Leu 370 375 380 LysLeu Phe Leu Ser Arg Arg Arg Trp Gln Arg Ser Gln Ile Ser Tyr 385 390 395400 Lys Cys His Leu Val Leu Arg Pro Ile Pro Glu Leu Leu Arg Ala Pro 405410 415 Pro Glu Arg Val Trp Val Thr Met Pro Arg Arg Pro Pro Pro Ser Gly420 425 430 Gly Leu Ser Ser Ser Ser Asp Ser Glu Glu Glu Glu Leu Glu GluLeu 435 440 445 Pro Ser Val Pro Arg Pro Leu Gln Pro Glu Phe Ser Gly SerArg Val 450 455 460 Ser Leu Thr Asp Ile Pro Thr Glu Asp Glu Ala Leu GlyThr Pro Glu 465 470 475 480 Thr Gly Ala Ala Thr Pro Met Asp Trp Gln GluGln Gly Arg Ala Pro 485 490 495 Ser Gln Asp Gln Glu Ala Pro Ser Pro GluAla Leu Pro Ser Pro Gly 500 505 510 Gln Glu Pro Ala Ala Gly Ala Ser ProArg Arg Gly Glu Leu Arg Arg 515 520 525 Gly Ser Ser Ala Glu Ser Ala LeuPro Arg Ala Gly Pro Arg Glu Leu 530 535 540 Gly Arg Gly Leu His Lys AlaAla Ser Val Glu Leu Pro Gln Arg Arg 545 550 555 560 Ser Pro Gly Pro GlyAla Thr Arg Leu Ala Arg Gly Gly Leu Gly Glu 565 570 575 Gly Glu Tyr AlaGln Arg Leu Gln Ala Leu Arg Gln Arg Leu Leu Arg 580 585 590 Gly Gly ProGlu Asp Gly Lys Val Ser Gly Leu Arg Gly Pro Leu Leu 595 600 605 Glu SerLeu Gly Gly Arg Ala Arg Asp Pro Arg Met Ala Arg Ala Ala 610 615 620 SerSer Glu Ala Ala Pro His His Gln Pro Pro Leu Glu Asn Arg Gly 625 630 635640 Leu Gln Lys Ser Ser Ser Phe Ser Gln Gly Glu Ala Glu Pro Arg Gly 645650 655 Arg His Arg Arg Ala Gly Ala Pro Leu Glu Ile Pro Val Ala Arg Leu660 665 670 Gly Ala Arg Arg Leu Gln Glu Ser Pro Ser Leu Ser Ala Leu SerGlu 675 680 685 Ala Gln Pro Ser Ser Pro Ala Arg Pro Ser Ala Pro Lys ProSer Thr 690 695 700 Pro Lys Ser Ala Glu Pro Ser Ala Thr Thr Pro Ser AspAla Pro Gln 705 710 715 720 Pro Pro Ala Pro Gln Pro Ala Gln Asp Lys AlaPro Glu Pro Arg Pro 725 730 735 Glu Pro Val Arg Ala Ser Lys Pro Ala ProPro Pro Gln Ala Leu Gln 740 745 750 Thr Leu Ala Leu Pro Leu Thr Pro TyrAla Gln Ile Ile Gln Ser Leu 755 760 765 Gln Leu Ser Gly His Ala Gln GlyPro Ser Gln Gly Pro Ala Ala Pro 770 775 780 Pro Ser Glu Pro Lys Pro HisAla Ala Val Phe Ala Arg Val Ala Ser 785 790 795 800 Pro Pro Pro Gly AlaPro Glu Lys Arg Val Pro Ser Ala Gly Gly Pro 805 810 815 Pro Val Leu AlaGlu Lys Ala Arg Val Pro Thr Val Pro Pro Arg Pro 820 825 830 Gly Ser SerLeu Ser Ser Ser Ile Glu Asn Leu Glu Ser Glu 835 840 845 5 279 PRT Homosapiens 5 Ser Pro Ala Lys Glu Val Val Ser Ser Pro Gly Ser Ser Pro ArgSer 1 5 10 15 Ser Pro Arg Pro Glu Gly Thr Thr Leu Arg Gln Gly Pro ProGln Lys 20 25 30 Pro Tyr Thr Phe Leu Glu Glu Lys Ala Arg Gly Arg Phe GlyVal Val 35 40 45 Arg Ala Cys Arg Glu Asn Ala Thr Gly Arg Thr Phe Val AlaLys Ile 50 55 60 Val Pro Tyr Ala Ala Glu Gly Lys Pro Arg Val Leu Gln GluTyr Glu 65 70 75 80 Val Leu Arg Thr Leu His His Glu Arg Ile Met Ser LeuHis Glu Ala 85 90 95 Tyr Ile Thr Pro Arg Tyr Leu Val Leu Ile Ala Glu SerCys Gly Asn 100 105 110 Arg Glu Leu Leu Cys Gly Leu Ser Asp Arg Phe ArgTyr Ser Glu Asp 115 120 125 Asp Val Ala Thr Tyr Met Val Gln Leu Leu GlnGly Leu Asp Tyr Leu 130 135 140 His Gly His His Val Leu His Leu Asp IleLys Pro Asp Asn Leu Leu 145 150 155 160 Leu Ala Pro Asp Asn Ala Leu LysIle Val Asp Phe Gly Ser Ala Gln 165 170 175 Pro Tyr Asn Pro Gln Ala LeuArg Pro Leu Gly His Arg Thr Gly Thr 180 185 190 Leu Glu Phe Met Ala ProGlu Met Val Lys Gly Glu Pro Ile Gly Ser 195 200 205 Ala Thr Asp Ile TrpGly Ala Gly Val Leu Thr Tyr Ile Met Leu Ser 210 215 220 Gly Arg Ser ProPhe Tyr Glu Pro Asp Pro Gln Glu Thr Glu Ala Arg 225 230 235 240 Ile ValGly Gly Arg Phe Asp Ala Phe Gln Leu Tyr Pro Asn Thr Ser 245 250 255 GlnSer Ala Thr Leu Phe Leu Arg Lys Val Leu Ser Val His Pro Trp 260 265 270Ser Arg Pro Ser Ser Cys Leu 275 6 549 PRT Homo sapiens 6 Leu Arg Glu ProGly Trp Ala Ala Thr Gly Leu Arg Lys Gly Val Gln 1 5 10 15 His Ile PheArg Val Leu Ser Thr Thr Val Lys Ser Ser Ser Lys Pro 20 25 30 Ser Pro ProSer Glu Pro Val Gln Leu Leu Glu His Gly Pro Thr Leu 35 40 45 Glu Glu AlaPro Ala Met Leu Asp Lys Pro Asp Ile Val Tyr Val Val 50 55 60 Glu Gly GlnPro Ala Ser Val Thr Val Thr Phe Asn His Val Glu Ala 65 70 75 80 Gln ValVal Trp Arg Ser Cys Arg Gly Ala Leu Leu Glu Ala Arg Ala 85 90 95 Gly ValTyr Glu Leu Ser Gln Pro Asp Asp Asp Gln Tyr Cys Leu Arg 100 105 110 IleCys Arg Val Ser Arg Arg Asp Met Gly Ala Leu Thr Cys Thr Ala 115 120 125Arg Asn Arg His Gly Thr Gln Thr Cys Ser Val Thr Leu Glu Leu Ala 130 135140 Glu Ala Pro Arg Phe Glu Ser Ile Met Glu Asp Val Glu Val Gly Ala 145150 155 160 Gly Glu Thr Ala Arg Phe Ala Val Val Val Glu Gly Lys Pro LeuPro 165 170 175 Asp Ile Met Trp Tyr Lys Asp Glu Val Leu Leu Thr Glu SerSer His 180 185 190 Val Ser Phe Val Tyr Glu Glu Asn Glu Cys Ser Leu ValVal Leu Ser 195 200 205 Thr Gly Ala Gln Asp Gly Gly Val Tyr Thr Cys ThrAla Gln Asn Leu 210 215 220 Ala Gly Glu Val Ser Cys Lys Ala Glu Leu AlaVal His Ser Ala Gln 225 230 235 240 Thr Ala Met Glu Val Glu Gly Val GlyGlu Asp Glu Asp His Arg Gly 245 250 255 Arg Arg Leu Ser Asp Phe Tyr AspIle His Gln Glu Ile Gly Arg Gly 260 265 270 Ala Phe Ser Tyr Leu Arg ArgIle Val Glu Arg Ser Ser Gly Leu Glu 275 280 285 Phe Ala Ala Lys Phe IlePro Ser Gln Ala Lys Pro Lys Ala Ser Ala 290 295 300 Arg Arg Glu Ala ArgLeu Leu Ala Arg Leu Gln His Asp Cys Val Leu 305 310 315 320 Tyr Phe HisGlu Ala Phe Glu Arg Arg Arg Gly Leu Val Ile Val Thr 325 330 335 Glu LeuCys Thr Glu Glu Leu Leu Glu Arg Ile Ala Arg Lys Pro Thr 340 345 350 ValCys Glu Ser Glu Ile Arg Ala Tyr Met Arg Gln Val Leu Glu Gly 355 360 365Ile His Tyr Leu His Gln Ser His Val Leu His Leu Asp Val Lys Pro 370 375380 Glu Asn Leu Leu Val Trp Asp Gly Ala Ala Gly Glu Gln Gln Val Arg 385390 395 400 Ile Cys Asp Phe Gly Asn Ala Gln Glu Leu Thr Pro Gly Glu ProGln 405 410 415 Tyr Cys Gln Tyr Gly Thr Pro Glu Phe Val Ala Pro Glu IleVal Asn 420 425 430 Gln Ser Pro Val Ser Gly Val Thr Asp Ile Trp Pro ValGly Val Val 435 440 445 Ala Phe Leu Cys Leu Thr Gly Ile Ser Pro Phe ValGly Glu Asn Asp 450 455 460 Arg Thr Thr Leu Met Asn Ile Arg Asn Tyr AsnVal Ala Phe Glu Glu 465 470 475 480 Thr Thr Phe Leu Ser Leu Ser Arg GluAla Arg Gly Phe Leu Ile Lys 485 490 495 Val Leu Val Gln Asp Arg Leu ArgPro Thr Ala Glu Glu Thr Leu Glu 500 505 510 His Pro Trp Phe Lys Thr GlnAla Lys Gly Ala Glu Val Ser Thr Asp 515 520 525 His Leu Lys Leu Phe LeuSer Arg Arg Arg Trp Gln Arg Ser Gln Ile 530 535 540 Ser Tyr Lys Cys His545 7 250 PRT Homo sapiens 7 Tyr Thr Phe Leu Glu Glu Lys Ala Arg Gly ArgPhe Gly Val Val Arg 1 5 10 15 Ala Cys Arg Glu Asn Ala Thr Gly Arg ThrPhe Val Ala Lys Ile Val 20 25 30 Pro Tyr Ala Ala Glu Gly Lys Pro Arg ValLeu Gln Glu Tyr Glu Val 35 40 45 Leu Arg Thr Leu His His Glu Arg Ile MetSer Leu His Glu Ala Tyr 50 55 60 Ile Thr Pro Arg Tyr Leu Val Leu Ile AlaGlu Ser Cys Gly Asn Arg 65 70 75 80 Glu Leu Leu Cys Gly Leu Ser Asp ArgPhe Arg Tyr Ser Glu Asp Asp 85 90 95 Val Ala Thr Tyr Met Val Gln Leu LeuGln Gly Leu Asp Tyr Leu His 100 105 110 Gly His His Val Leu His Leu AspIle Lys Pro Asp Asn Leu Leu Leu 115 120 125 Ala Pro Asp Asn Ala Leu LysIle Val Asp Phe Gly Ser Ala Gln Pro 130 135 140 Tyr Asn Pro Gln Ala LeuArg Pro Leu Gly His Arg Thr Gly Thr Leu 145 150 155 160 Glu Phe Met AlaPro Glu Met Val Lys Gly Glu Pro Ile Gly Ser Ala 165 170 175 Thr Asp IleTrp Gly Ala Gly Val Leu Thr Tyr Ile Met Leu Ser Gly 180 185 190 Arg SerPro Phe Tyr Glu Pro Asp Pro Gln Glu Thr Glu Ala Arg Ile 195 200 205 ValGly Gly Arg Phe Asp Ala Phe Gln Leu Tyr Pro Asn Thr Ser Gln 210 215 220Ser Ala Thr Leu Phe Leu Arg Lys Val Leu Ser Val His Pro Trp Ser 225 230235 240 Arg Pro Ser Ser Cys Leu Ser Val Cys His 245 250 8 245 PRT Homosapiens 8 Pro Arg Lys Asp Lys Gly Leu Ser Pro Pro Asn Leu Ser Ala SerVal 1 5 10 15 Gln Glu Glu Leu Gly His Gln Tyr Val Arg Ser Glu Ser AspPhe Pro 20 25 30 Pro Val Phe His Ile Lys Leu Lys Asp Gln Val Leu Leu GluGly Glu 35 40 45 Ala Ala Thr Leu Leu Cys Leu Pro Ala Ala Cys Pro Ala ProHis Ile 50 55 60 Ser Trp Met Lys Asp Lys Lys Ser Leu Arg Ser Glu Pro SerVal Ile 65 70 75 80 Ile Val Ser Cys Lys Asp Gly Arg Gln Leu Leu Ser IlePro Arg Ala 85 90 95 Gly Lys Arg His Ala Gly Leu Tyr Glu Cys Ser Ala ThrAsn Val Leu 100 105 110 Gly Ser Ile Thr Ser Ser Cys Thr Val Ala Val AlaArg Val Pro Gly 115 120 125 Lys Leu Ala Pro Pro Glu Val Thr Gln Thr TyrGln Asp Thr Ala Leu 130 135 140 Val Leu Trp Lys Pro Gly Asp Ser Arg AlaPro Cys Thr Tyr Thr Leu 145 150 155 160 Glu Arg Arg Val Asp Gly Glu SerVal Trp His Pro Val Ser Ser Gly 165 170 175 Ile Pro Asp Cys Tyr Tyr AsnVal Thr His Leu Pro Val Gly Val Thr 180 185 190 Val Arg Phe Arg Val AlaCys Ala Asn Arg Ala Gly Gln Gly Pro Phe 195 200 205 Ser Asn Ser Ser GluLys Val Phe Val Arg Gly Thr Gln Asp Ser Ser 210 215 220 Ala Val Pro SerAla Ala His Gln Glu Ala Pro Val Thr Ser Arg Pro 225 230 235 240 Ala ArgAla Arg Pro 245 9 111 PRT Homo sapiens 9 Leu Glu Asp Val Glu Val Leu GluGly Arg Ala Ala Arg Phe Asp Cys 1 5 10 15 Lys Ile Ser Gly Thr Pro ProPro Val Val Thr Trp Thr His Phe Gly 20 25 30 Cys Pro Met Glu Glu Ser GluAsn Leu Arg Leu Arg Gln Asp Gly Gly 35 40 45 Leu His Ser Leu His Ile AlaHis Val Gly Ser Glu Asp Glu Gly Leu 50 55 60 Tyr Ala Val Ser Ala Val AsnThr His Gly Gln Ala His Cys Ser Ala 65 70 75 80 Gln Leu Tyr Val Glu GluPro Arg Thr Ala Ala Ser Gly Pro Ser Ser 85 90 95 Lys Leu Glu Lys Met ProSer Ile Pro Glu Glu Pro Glu Gln Gly 100 105 110 10 198 PRT Homo sapiens10 Pro Asp Phe Leu Arg Pro Leu Gln Asp Leu Glu Val Gly Leu Ala Lys 1 510 15 Glu Ala Met Leu Glu Cys Gln Val Thr Gly Leu Pro Tyr Pro Thr Ile 2025 30 Ser Trp Phe His Asn Gly His Arg Ile Gln Ser Ser Asp Asp Arg Arg 3540 45 Met Thr Gln Tyr Arg Asp Val His Arg Leu Val Phe Pro Ala Val Gly 5055 60 Pro Gln His Ala Gly Val Tyr Lys Ser Val Ile Ala Asn Lys Leu Gly 6570 75 80 Lys Ala Ala Cys Tyr Ala His Leu Tyr Val Thr Asp Val Val Pro Gly85 90 95 Pro Pro Asp Gly Ala Pro Gln Val Val Ala Val Thr Gly Arg Met Val100 105 110 Thr Leu Thr Trp Asn Pro Pro Arg Ser Leu Asp Met Ala Ile AspPro 115 120 125 Asp Ser Leu Thr Tyr Thr Val Gln His Gln Val Leu Gly SerAsp Gln 130 135 140 Trp Thr Ala Leu Val Thr Gly Leu Arg Glu Pro Gly TrpAla Ala Thr 145 150 155 160 Gly Leu Arg Lys Gly Val Gln His Ile Phe ArgVal Leu Ser Thr Thr 165 170 175 Val Lys Ser Ser Ser Lys Pro Ser Pro ProSer Glu Pro Val Gln Leu 180 185 190 Leu Glu His Gly Pro Thr 195 11 101PRT Homo sapiens 11 Ala Pro Leu Phe Thr Arg Leu Leu Glu Asp Val Glu ValLeu Glu Gly 1 5 10 15 Arg Ala Ala Arg Phe Asp Cys Lys Ile Ser Gly ThrPro Pro Pro Val 20 25 30 Val Thr Trp Thr His Phe Gly Cys Pro Met Glu GluSer Glu Asn Leu 35 40 45 Arg Leu Arg Gln Asp Gly Gly Leu His Ser Leu HisIle Ala His Val 50 55 60 Gly Ser Glu Asp Glu Gly Leu Tyr Ala Val Ser AlaVal Asn Thr His 65 70 75 80 Gly Gln Ala His Cys Ser Ala Gln Leu Tyr ValGlu Glu Pro Arg Thr 85 90 95 Ala Ala Ser Gly Pro 100 12 195 PRT Homosapiens 12 Arg Gly Thr Gln Asp Ser Ser Ala Val Pro Ser Ala Ala His GlnGlu 1 5 10 15 Ala Pro Val Thr Ser Arg Pro Ala Arg Ala Arg Pro Pro AspSer Pro 20 25 30 Thr Ser Leu Ala Pro Pro Leu Ala Pro Ala Ala Pro Thr ProPro Ser 35 40 45 Val Thr Val Ser Pro Ser Ser Pro Pro Thr Pro Pro Ser GlnAla Leu 50 55 60 Ser Ser Leu Lys Ala Val Gly Pro Pro Pro Gln Thr Pro ProArg Arg 65 70 75 80 His Arg Gly Leu Gln Ala Ala Arg Pro Ala Glu Pro ThrLeu Pro Ser 85 90 95 Thr His Val Thr Pro Ser Glu Pro Lys Pro Phe Val LeuAsp Thr Gly 100 105 110 Thr Pro Ile Pro Ala Ser Thr Pro Gln Gly Val LysPro Val Ser Ser 115 120 125 Ser Thr Pro Val Tyr Val Val Thr Ser Phe ValSer Ala Pro Pro Ala 130 135 140 Pro Glu Pro Pro Ala Pro Glu Pro Pro ProGlu Pro Thr Lys Val Thr 145 150 155 160 Val Gln Ser Leu Ser Pro Ala LysGlu Val Val Ser Ser Pro Gly Ser 165 170 175 Ser Pro Arg Ser Ser Pro ArgPro Glu Gly Thr Thr Leu Arg Gln Gly 180 185 190 Pro Pro Gln 195 13 90PRT Homo sapiens 13 Pro Asp Phe Leu Arg Pro Leu Gln Asp Leu Glu Val GlyLeu Ala Lys 1 5 10 15 Glu Ala Met Leu Glu Cys Gln Val Thr Gly Leu ProTyr Pro Thr Ile 20 25 30 Ser Trp Phe His Asn Gly His Arg Ile Gln Ser SerAsp Asp Arg Arg 35 40 45 Met Thr Gln Tyr Arg Asp Val His Arg Leu Val PhePro Ala Val Gly 50 55 60 Pro Gln His Ala Gly Val Tyr Lys Ser Val Ile AlaAsn Lys Leu Gly 65 70 75 80 Lys Ala Ala Cys Tyr Ala His Leu Tyr Val 8590 14 414 PRT Homo sapiens 14 Pro Pro Glu Phe Val Ile Pro Leu Ser GluVal Thr Cys Glu Thr Gly 1 5 10 15 Glu Thr Val Val Leu Arg Cys Arg ValCys Gly Arg Pro Lys Ala Ser 20 25 30 Ile Thr Trp Lys Gly Pro Glu His AsnThr Leu Asn Asn Asp Gly His 35 40 45 Tyr Ser Ile Ser Tyr Ser Asp Leu GlyGlu Ala Thr Leu Lys Ile Val 50 55 60 Gly Val Thr Thr Glu Asp Asp Gly IleTyr Thr Cys Ile Ala Val Asn 65 70 75 80 Asp Met Gly Ser Ala Ser Ser SerAla Ser Leu Arg Val Leu Gly Pro 85 90 95 Gly Met Asp Gly Ile Met Val ThrTrp Lys Asp Asn Phe Asp Ser Phe 100 105 110 Tyr Ser Glu Val Ala Glu LeuGly Arg Gly Arg Phe Ser Val Val Lys 115 120 125 Lys Cys Asp Gln Lys GlyThr Lys Arg Ala Val Ala Thr Lys Phe Val 130 135 140 Asn Lys Lys Leu MetLys Arg Asp Gln Val Thr His Glu Leu Gly Ile 145 150 155 160 Leu Gln SerLeu Gln His Pro Leu Leu Val Gly Leu Leu Asp Thr Phe 165 170 175 Glu ThrPro Thr Ser Tyr Ile Leu Val Leu Glu Met Ala Asp Gln Gly 180 185 190 ArgLeu Leu Asp Cys Val Val Arg Trp Gly Ser Leu Thr Glu Gly Lys 195 200 205Ile Arg Ala His Leu Gly Glu Val Leu Glu Ala Val Arg Tyr Leu His 210 215220 Asn Cys Arg Ile Ala His Leu Asp Leu Lys Pro Glu Asn Ile Leu Val 225230 235 240 Asp Glu Ser Leu Ala Lys Pro Thr Ile Lys Leu Ala Asp Phe GlyAsp 245 250 255 Ala Val Gln Leu Asn Thr Thr Tyr Tyr Ile His Gln Leu LeuGly Asn 260 265 270 Pro Glu Phe Ala Ala Pro Glu Ile Ile Leu Gly Asn ProVal Ser Leu 275 280 285 Thr Ser Asp Thr Trp Ser Val Gly Val Leu Thr TyrVal Leu Leu Ser 290 295 300 Gly Val Ser Pro Phe Leu Asp Asp Ser Val GluGlu Thr Cys Leu Asn 305 310 315 320 Ile Cys Arg Leu Asp Phe Ser Phe ProAsp Asp Tyr Phe Lys Gly Val 325 330 335 Ser Gln Lys Ala Lys Glu Phe ValCys Phe Leu Leu Gln Glu Asp Pro 340 345 350 Ala Lys Arg Pro Ser Ala AlaLeu Ala Leu Gln Glu Gln Trp Leu Gln 355 360 365 Ala Gly Asn Gly Arg SerThr Gly Val Leu Asp Thr Ser Arg Leu Thr 370 375 380 Ser Phe Ile Glu ArgArg Lys His Gln Asn Asp Val Arg Pro Ile Arg 385 390 395 400 Ser Ile LysAsn Phe Leu Gln Ser Arg Leu Leu Pro Arg Val 405 410 15 274 PRT Homosapiens 15 Glu Leu Gly Arg Gly Arg Phe Ser Val Val Lys Lys Cys Asp GlnLys 1 5 10 15 Gly Thr Lys Arg Ala Val Ala Thr Lys Phe Val Asn Lys LysLeu Met 20 25 30 Lys Arg Asp Gln Val Thr His Glu Leu Gly Ile Leu Gln SerLeu Gln 35 40 45 His Pro Leu Leu Val Gly Leu Leu Asp Thr Phe Glu Thr ProThr Ser 50 55 60 Tyr Ile Leu Val Leu Glu Met Ala Asp Gln Gly Arg Leu LeuAsp Cys 65 70 75 80 Val Val Arg Trp Gly Ser Leu Thr Glu Gly Lys Ile ArgAla His Leu 85 90 95 Gly Glu Val Leu Glu Ala Val Arg Tyr Leu His Asn CysArg Ile Ala 100 105 110 His Leu Asp Leu Lys Pro Glu Asn Ile Leu Val AspGlu Ser Leu Ala 115 120 125 Lys Pro Thr Ile Lys Leu Ala Asp Phe Gly AspAla Val Gln Leu Asn 130 135 140 Thr Thr Tyr Tyr Ile His Gln Leu Leu GlyAsn Pro Glu Phe Ala Ala 145 150 155 160 Pro Glu Ile Ile Leu Gly Asn ProVal Ser Leu Thr Ser Asp Thr Trp 165 170 175 Ser Val Gly Val Leu Thr TyrVal Leu Leu Ser Gly Val Ser Pro Phe 180 185 190 Leu Asp Asp Ser Val GluGlu Thr Cys Leu Asn Ile Cys Arg Leu Asp 195 200 205 Phe Ser Phe Pro AspAsp Tyr Phe Lys Gly Val Ser Gln Lys Ala Lys 210 215 220 Glu Phe Val CysPhe Leu Leu Gln Glu Asp Pro Ala Lys Arg Pro Ser 225 230 235 240 Ala AlaLeu Ala Leu Gln Glu Gln Trp Leu Gln Ala Gly Asn Gly Arg 245 250 255 SerThr Gly Val Leu Asp Thr Ser Arg Leu Thr Ser Phe Ile Glu Arg 260 265 270Arg Lys 16 141 PRT Homo sapiens 16 Gly Lys Arg Glu Gly Lys Leu Glu AsnGly Tyr Arg Lys Ser Arg Glu 1 5 10 15 Gly Leu Ser Asn Lys Val Ser ValLys Leu Leu Asn Pro Asn Tyr Ile 20 25 30 Tyr Asp Val Pro Pro Glu Phe ValIle Pro Leu Ser Glu Val Thr Cys 35 40 45 Glu Thr Gly Glu Thr Val Val LeuArg Cys Arg Val Cys Gly Arg Pro 50 55 60 Lys Ala Ser Ile Thr Trp Lys GlyPro Glu His Asn Thr Leu Asn Asn 65 70 75 80 Asp Gly His Tyr Ser Ile SerTyr Ser Asp Leu Gly Glu Ala Thr Leu 85 90 95 Lys Ile Val Gly Val Thr ThrGlu Asp Asp Gly Ile Tyr Thr Cys Ile 100 105 110 Ala Val Asn Asp Met GlySer Ala Ser Ser Ser Ala Ser Leu Arg Val 115 120 125 Leu Gly Pro Gly MetAsp Gly Ile Met Val Thr Trp Lys 130 135 140 17 196 PRT Homo sapiens 17Gly Gly Ala Pro Ser Gly Gly Ser Gly His Ser Gly Gly Pro Ser Ser 1 5 1015 Cys Gly Gly Ala Pro Ser Thr Ser Arg Ser Arg Pro Ser Arg Ile Pro 20 2530 Gln Pro Val Arg His His Pro Pro Val Leu Val Ser Ser Ala Ala Ser 35 4045 Ser Gln Ala Glu Ala Asp Lys Met Ser Gly Thr Ser Thr Pro Gly Pro 50 5560 Ser Leu Pro Pro Pro Gly Ala Ala Pro Glu Ala Gly Pro Ser Ala Pro 65 7075 80 Ser Arg Arg Pro Pro Gly Ala Asp Ala Glu Gly Ser Glu Arg Glu Ala 8590 95 Glu Pro Ile Pro Lys Met Lys Val Leu Glu Ser Pro Arg Lys Gly Ala100 105 110 Ala Asn Ala Ser Gly Ser Ser Pro Asp Ala Pro Ala Lys Asp AlaArg 115 120 125 Ala Ser Leu Gly Thr Leu Pro Leu Gly Lys Pro Arg Ala GlyAla Ala 130 135 140 Ser Pro Leu Asn Ser Pro Leu Ser Ser Ala Val Pro SerLeu Gly Lys 145 150 155 160 Glu Pro Phe Pro Pro Ser Ser Pro Leu Gln LysGly Gly Ser Phe Trp 165 170 175 Ser Ser Ile Pro Ala Ser Pro Ala Ser ArgPro Gly Ser Phe Thr Phe 180 185 190 Pro Gly Asp Ser 195 18 298 PRT Homosapiens 18 Gln Lys Val Ser Asp Phe Tyr Asp Ile Glu Glu Arg Leu Gly SerGly 1 5 10 15 Lys Phe Gly Gln Val Phe Arg Leu Val Glu Lys Lys Thr ArgLys Val 20 25 30 Trp Ala Gly Lys Phe Phe Lys Ala Tyr Ser Ala Lys Glu LysGlu Asn 35 40 45 Ile Arg Gln Glu Ile Ser Ile Met Asn Cys Leu His His ProLys Leu 50 55 60 Val Gln Cys Val Asp Ala Phe Glu Glu Lys Ala Asn Ile ValMet Val 65 70 75 80 Leu Glu Ile Val Ser Gly Gly Glu Leu Phe Glu Arg IleIle Asp Glu 85 90 95 Asp Phe Glu Leu Thr Glu Arg Glu Cys Ile Lys Tyr MetArg Gln Ile 100 105 110 Ser Glu Gly Val Glu Tyr Ile His Lys Gln Gly IleVal His Leu Asp 115 120 125 Leu Lys Pro Glu Asn Ile Met Cys Val Asn LysThr Gly Thr Arg Ile 130 135 140 Lys Leu Ile Asp Phe Gly Leu Ala Arg ArgLeu Glu Asn Ala Gly Ser 145 150 155 160 Leu Lys Val Leu Phe Gly Thr ProGlu Phe Val Ala Pro Glu Val Ile 165 170 175 Asn Tyr Glu Pro Ile Ser TyrAla Thr Asp Met Trp Ser Ile Gly Val 180 185 190 Ile Cys Tyr Ile Leu ValSer Gly Leu Ser Pro Phe Met Gly Asp Asn 195 200 205 Asp Asn Glu Thr LeuAla Asn Val Thr Ser Ala Thr Trp Asp Phe Asp 210 215 220 Asp Glu Ala PheAsp Glu Ile Ser Asp Asp Ala Lys Asp Phe Ile Ser 225 230 235 240 Asn LeuLeu Lys Lys Asp Met Lys Asn Arg Leu Asp Cys Thr Gln Cys 245 250 255 LeuGln His Pro Trp Leu Met Lys Asp Thr Lys Asn Met Glu Ala Lys 260 265 270Lys Leu Ser Lys Asp Arg Met Lys Lys Tyr Met Ala Arg Arg Lys Trp 275 280285 Gln Lys Thr Gly Asn Ala Val Arg Ala Ile 290 295 19 508 PRT Homosapiens 19 Gly Thr Glu Ser Asp Ala Thr Val Lys Lys Lys Pro Ala Pro LysThr 1 5 10 15 Pro Pro Lys Ala Ala Met Pro Pro Gln Ile Ile Gln Phe ProGlu Asp 20 25 30 Gln Lys Val Arg Ala Gly Glu Ser Val Glu Leu Phe Gly LysVal Thr 35 40 45 Gly Thr Gln Pro Ile Thr Cys Thr Trp Met Lys Phe Arg LysGln Ile 50 55 60 Gln Asp Ser Glu His Ile Lys Val Glu Asn Ser Glu Asn GlySer Lys 65 70 75 80 Leu Thr Ile Leu Ala Ala Arg Gln Glu His Cys Gly CysTyr Thr Leu 85 90 95 Leu Val Glu Asn Lys Leu Gly Ser Arg Gln Ala Gln ValAsn Leu Thr 100 105 110 Val Val Asp Lys Pro Asp Pro Pro Ala Gly Thr ProCys Ala Ser Asp 115 120 125 Ile Arg Ser Ser Ser Leu Thr Leu Ser Trp TyrGly Ser Ser Tyr Asp 130 135 140 Gly Gly Ser Ala Val Gln Ser Tyr Ser IleGlu Ile Trp Asp Ser Ala 145 150 155 160 Asn Lys Thr Trp Lys Glu Leu AlaThr Cys Arg Ser Thr Ser Phe Asn 165 170 175 Val Gln Asp Leu Leu Pro AspHis Glu Tyr Lys Phe Arg Val Arg Ala 180 185 190 Ile Asn Val Tyr Gly ThrSer Glu Pro Ser Gln Glu Ser Glu Leu Thr 195 200 205 Thr Val Gly Glu LysPro Glu Glu Pro Lys Met Lys Trp Arg Cys Gln 210 215 220 Thr Asp Asp GluLys Glu Pro Glu Val Asp Tyr Arg Thr Val Thr Ile 225 230 235 240 Asn ThrGlu Gln Lys Val Ser Asp Phe Tyr Asp Ile Glu Glu Arg Leu 245 250 255 GlySer Gly Lys Phe Gly Gln Val Phe Arg Leu Val Glu Lys Lys Thr 260 265 270Arg Lys Val Trp Ala Gly Lys Phe Phe Lys Ala Tyr Ser Ala Lys Glu 275 280285 Lys Glu Asn Ile Arg Gln Glu Ile Ser Ile Met Asn Cys Leu His His 290295 300 Pro Lys Leu Val Gln Cys Val Asp Ala Phe Glu Glu Lys Ala Asn Ile305 310 315 320 Val Met Val Leu Glu Ile Val Ser Gly Gly Glu Leu Phe GluArg Ile 325 330 335 Ile Asp Glu Asp Phe Glu Leu Thr Glu Arg Glu Cys IleLys Tyr Met 340 345 350 Arg Gln Ile Ser Glu Gly Val Glu Tyr Ile His LysGln Gly Ile Val 355 360 365 His Leu Asp Leu Lys Pro Glu Asn Ile Met CysVal Asn Lys Thr Gly 370 375 380 Thr Arg Ile Lys Leu Ile Asp Phe Gly LeuAla Arg Arg Leu Glu Asn 385 390 395 400 Ala Gly Ser Leu Lys Val Leu PheGly Thr Pro Glu Phe Val Ala Pro 405 410 415 Glu Val Ile Asn Tyr Glu ProIle Ser Tyr Ala Thr Asp Met Trp Ser 420 425 430 Ile Gly Val Ile Cys TyrIle Leu Val Ser Gly Leu Ser Pro Phe Met 435 440 445 Gly Asp Asn Asp AsnGlu Thr Leu Ala Asn Val Thr Ser Ala Thr Trp 450 455 460 Asp Phe Asp AspGlu Ala Phe Asp Glu Ile Ser Asp Asp Ala Lys Asp 465 470 475 480 Phe IleSer Asn Leu Leu Lys Lys Asp Met Lys Asn Arg Leu Asp Cys 485 490 495 ThrGln Cys Leu Gln His Pro Trp Leu Met Lys Asp 500 505 20 106 PRT Homosapiens 20 Pro Tyr Phe Ser Lys Thr Ile Arg Asp Leu Glu Val Val Glu GlySer 1 5 10 15 Ala Ala Arg Phe Asp Cys Lys Ile Glu Gly Tyr Pro Asp ProGlu Val 20 25 30 Val Trp Phe Lys Asp Asp Gln Ser Ile Arg Glu Ser Arg HisPhe Gln 35 40 45 Ile Asp Tyr Asp Glu Asp Gly Asn Cys Ser Leu Ile Ile SerAsp Val 50 55 60 Cys Gly Asp Asp Asp Ala Lys Tyr Thr Cys Lys Ala Val AsnSer Leu 65 70 75 80 Gly Glu Ala Thr Cys Thr Ala Glu Leu Ile Val Glu ThrMet Glu Glu 85 90 95 Gly Glu Gly Glu Gly Glu Glu Glu Glu Glu 100 105 2196 PRT Homo sapiens 21 Pro Pro Lys Phe Ala Thr Lys Leu Gly Arg Val ValVal Lys Glu Gly 1 5 10 15 Gln Met Gly Arg Phe Ser Cys Lys Ile Thr GlyArg Pro Gln Pro Gln 20 25 30 Val Thr Trp Leu Lys Gly Asn Val Pro Leu GlnPro Ser Ala Arg Val 35 40 45 Ser Val Ser Glu Lys Asn Gly Met Gln Val LeuGlu Ile His Gly Val 50 55 60 Asn Gln Asp Asp Val Gly Val Tyr Thr Cys LeuVal Val Asn Gly Ser 65 70 75 80 Gly Lys Ala Ser Met Ser Ala Glu Leu SerIle Gln Gly Leu Asp Ser 85 90 95 22 96 PRT Homo sapiens 22 Pro Pro LysPhe Ala Thr Lys Leu Gly Arg Val Val Val Lys Glu Gly 1 5 10 15 Gln MetGly Arg Phe Ser Cys Lys Ile Thr Gly Arg Pro Gln Pro Gln 20 25 30 Val ThrTrp Leu Lys Gly Asn Val Pro Leu Gln Pro Ser Ala Arg Val 35 40 45 Ser ValSer Glu Lys Asn Gly Met Gln Val Leu Glu Ile His Gly Val 50 55 60 Asn GlnAsp Asp Val Gly Val Tyr Thr Cys Leu Val Val Asn Gly Ser 65 70 75 80 GlyLys Ala Ser Met Ser Ala Glu Leu Ser Ile Gln Gly Leu Asp Ser 85 90 95 2388 PRT Homo sapiens 23 Pro Lys Phe Ala Thr Lys Leu Gly Arg Val Val ValLys Glu Gly Gln 1 5 10 15 Met Gly Arg Phe Ser Cys Lys Ile Thr Gly ArgPro Gln Pro Gln Val 20 25 30 Thr Trp Leu Lys Gly Asn Val Pro Leu Gln ProSer Ala Arg Val Ser 35 40 45 Val Ser Glu Lys Asn Gly Met Gln Val Leu GluIle His Gly Val Asn 50 55 60 Gln Asp Asp Val Gly Val Tyr Thr Cys Leu ValVal Asn Gly Ser Gly 65 70 75 80 Lys Ala Ser Met Ser Ala Glu Leu 85 24 94PRT Homo sapiens 24 Ala Pro Ser Phe Ser Ser Val Leu Lys Asp Cys Ala ValIle Glu Gly 1 5 10 15 Gln Asp Phe Val Leu Gln Cys Ser Val Arg Gly ThrPro Val Pro Arg 20 25 30 Ile Thr Trp Leu Leu Asn Gly Gln Pro Ile Gln TyrAla Arg Ser Thr 35 40 45 Cys Glu Ala Gly Val Ala Glu Leu His Ile Gln AspAla Leu Pro Glu 50 55 60 Asp His Gly Thr Tyr Thr Cys Leu Ala Glu Asn AlaLeu Gly Gln Val 65 70 75 80 Ser Cys Ser Ala Trp Val Thr Val His Glu LysLys Ser Ser 85 90 25 112 PRT Homo sapiens 25 Lys Lys Ser Ser Arg Lys SerGlu Tyr Leu Leu Pro Val Ala Pro Ser 1 5 10 15 Lys Pro Thr Ala Pro IlePhe Leu Gln Gly Leu Ser Asp Leu Lys Val 20 25 30 Met Asp Gly Ser Gln ValThr Met Thr Val Gln Val Ser Gly Asn Pro 35 40 45 Pro Pro Glu Val Ile TrpLeu His Asn Gly Asn Glu Ile Gln Glu Ser 50 55 60 Glu Asp Phe His Phe GluGln Arg Gly Thr Gln His Ser Leu Trp Ile 65 70 75 80 Gln Glu Val Phe ProGlu Asp Thr Gly Thr Tyr Thr Cys Glu Ala Trp 85 90 95 Asn Ser Ala Gly GluVal Arg Thr Gln Ala Val Leu Thr Val Gln Glu 100 105 110 26 100 PRT Homosapiens 26 Ser Met Pro Leu Thr Glu Ala Pro Ala Phe Ile Leu Pro Pro ArgAsn 1 5 10 15 Leu Cys Ile Lys Glu Gly Ala Thr Ala Lys Phe Glu Gly ArgVal Arg 20 25 30 Gly Tyr Pro Glu Pro Gln Val Thr Trp His Arg Asn Gly GlnPro Ile 35 40 45 Thr Ser Gly Gly Arg Phe Leu Leu Asp Cys Gly Ile Arg GlyThr Phe 50 55 60 Ser Leu Val Ile His Ala Val His Glu Glu Asp Arg Gly LysTyr Thr 65 70 75 80 Cys Glu Ala Thr Asn Gly Ser Gly Ala Arg Gln Val ThrVal Glu Leu 85 90 95 Thr Val Glu Gly 100 27 174 PRT Homo sapiens 27 ProSer Gly Glu Glu Arg Lys Arg Pro Ala Pro Pro Arg Pro Ala Thr 1 5 10 15Phe Pro Thr Arg Gln Pro Gly Leu Gly Ser Gln Asp Val Val Ser Lys 20 25 30Ala Ala Asn Arg Arg Ile Pro Met Glu Gly Gln Arg Asp Ser Ala Phe 35 40 45Pro Lys Phe Glu Ser Lys Pro Gln Ser Gln Glu Val Lys Glu Asn Gln 50 55 60Thr Val Lys Phe Arg Cys Glu Val Ser Gly Ile Pro Lys Pro Glu Val 65 70 7580 Ala Trp Phe Leu Glu Gly Thr Pro Val Arg Arg Gln Glu Gly Ser Ile 85 9095 Glu Val Tyr Glu Asp Ala Gly Ser His Tyr Leu Cys Leu Leu Lys Ala 100105 110 Arg Thr Arg Asp Ser Gly Thr Tyr Ser Cys Thr Ala Ser Asn Ala Gln115 120 125 Gly Gln Val Ser Cys Ser Trp Thr Leu Gln Val Glu Arg Leu AlaVal 130 135 140 Met Glu Val Ala Pro Ser Phe Ser Ser Val Leu Lys Asp CysAla Val 145 150 155 160 Ile Glu Gly Gln Asp Phe Val Leu Gln Cys Ser ValArg Gly 165 170 28 97 PRT Homo sapiens 28 Pro Ala Phe Lys Gln Lys LeuGln Asp Val His Val Ala Glu Gly Lys 1 5 10 15 Lys Leu Leu Leu Gln CysGln Val Ser Ser Asp Pro Pro Ala Thr Ile 20 25 30 Ile Trp Thr Leu Asn GlyLys Thr Leu Lys Thr Thr Lys Phe Ile Ile 35 40 45 Leu Ser Gln Glu Gly SerLeu Cys Ser Val Ser Ile Glu Lys Ala Leu 50 55 60 Leu Glu Asp Arg Gly LeuTyr Lys Cys Val Ala Lys Asn Asp Ala Gly 65 70 75 80 Gln Ala Glu Cys SerCys Gln Val Thr Val Asp Asp Ala Pro Ala Ser 85 90 95 Glu 29 124 PRT Homosapiens 29 Glu Ser Gln Gly Thr Ala Pro Ala Phe Lys Gln Lys Leu Gln AspVal 1 5 10 15 His Val Ala Glu Gly Lys Lys Leu Leu Leu Gln Cys Gln ValSer Ser 20 25 30 Asp Pro Pro Ala Thr Ile Ile Trp Thr Leu Asn Gly Lys ThrLeu Lys 35 40 45 Thr Thr Lys Phe Ile Ile Leu Ser Gln Glu Gly Ser Leu CysSer Val 50 55 60 Ser Ile Glu Lys Ala Leu Leu Glu Asp Arg Gly Leu Tyr LysCys Val 65 70 75 80 Ala Lys Asn Asp Ala Gly Gln Ala Glu Cys Ser Cys GlnVal Thr Val 85 90 95 Asp Asp Ala Pro Ala Ser Glu Asn Thr Lys Ala Pro GluMet Lys Ser 100 105 110 Arg Arg Pro Lys Ser Ser Leu Pro Pro Val Leu Gly115 120 30 87 PRT Homo sapiens 30 Ala Pro Ala Phe Ile Leu Pro Pro ArgAsn Leu Cys Ile Lys Glu Gly 1 5 10 15 Ala Thr Ala Lys Phe Glu Gly ArgVal Arg Gly Tyr Pro Glu Pro Gln 20 25 30 Val Thr Trp His Arg Asn Gly GlnPro Ile Thr Ser Gly Gly Arg Phe 35 40 45 Leu Leu Asp Cys Gly Ile Arg GlyThr Phe Ser Leu Val Ile His Ala 50 55 60 Val His Glu Glu Asp Arg Gly LysTyr Thr Cys Glu Ala Thr Asn Gly 65 70 75 80 Ser Gly Ala Arg Gln Val Thr85 31 119 PRT Homo sapiens 31 Ser Asn Ala Gln Gly Gln Val Ser Cys SerTrp Thr Leu Gln Val Glu 1 5 10 15 Arg Leu Ala Val Met Glu Val Ala ProSer Phe Ser Ser Val Leu Lys 20 25 30 Asp Cys Ala Val Ile Glu Gly Gln AspPhe Val Leu Gln Cys Ser Val 35 40 45 Arg Gly Thr Pro Val Pro Arg Ile ThrTrp Leu Leu Asn Gly Gln Pro 50 55 60 Ile Gln Tyr Ala Arg Ser Thr Cys GluAla Gly Val Ala Glu Leu His 65 70 75 80 Ile Gln Asp Ala Leu Pro Glu AspHis Gly Thr Tyr Thr Cys Leu Ala 85 90 95 Glu Asn Ala Leu Gly Gln Val SerCys Ser Ala Trp Val Thr Val His 100 105 110 Glu Lys Lys Ser Ser Arg Lys115 32 98 PRT Homo sapiens 32 Gly Gln Arg Asp Ser Ala Phe Pro Lys PheGlu Ser Lys Pro Gln Ser 1 5 10 15 Gln Glu Val Lys Glu Asn Gln Thr ValLys Phe Arg Cys Glu Val Ser 20 25 30 Gly Ile Pro Lys Pro Glu Val Ala TrpPhe Leu Glu Gly Thr Pro Val 35 40 45 Arg Arg Gln Glu Gly Ser Ile Glu ValTyr Glu Asp Ala Gly Ser His 50 55 60 Tyr Leu Cys Leu Leu Lys Ala Arg ThrArg Asp Ser Gly Thr Tyr Ser 65 70 75 80 Cys Thr Ala Ser Asn Ala Gln GlyGln Val Ser Cys Ser Trp Thr Leu 85 90 95 Gln Val 33 82 PRT Homo sapiens33 Val Thr Ala Ser Leu Gly Gln Ser Val Leu Ile Ser Cys Ala Ile Ala 1 510 15 Gly Asp Pro Phe Pro Thr Val His Trp Leu Arg Asp Gly Lys Ala Leu 2025 30 Cys Lys Asp Thr Gly His Phe Glu Val Leu Gln Asn Glu Asp Val Phe 3540 45 Thr Leu Val Leu Lys Lys Val Gln Pro Trp His Ala Gly Gln Tyr Glu 5055 60 Ile Leu Leu Lys Asn Arg Val Gly Glu Cys Ser Cys Gln Val Ser Leu 6570 75 80 Met Leu 34 89 PRT Homo sapiens 34 Pro Tyr Phe Ser Lys Thr IleArg Asp Leu Glu Val Val Glu Gly Ser 1 5 10 15 Ala Ala Arg Phe Asp CysLys Ile Glu Gly Tyr Pro Asp Pro Glu Val 20 25 30 Val Trp Phe Lys Asp AspGln Ser Ile Arg Glu Ser Arg His Phe Gln 35 40 45 Ile Asp Tyr Asp Glu AspGly Asn Cys Ser Leu Ile Ile Ser Asp Val 50 55 60 Cys Gly Asp Asp Asp AlaLys Tyr Thr Cys Lys Ala Val Asn Ser Leu 65 70 75 80 Gly Glu Ala Thr CysThr Ala Glu Leu 85

That which is claimed is:
 1. An isolated peptide consisting of an aminoacid sequence selected from the group consisting of: (a) an amino acidsequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelicvariant of an amino acid sequence shown in SEQ ID NO:2, wherein saidallelic variant is encoded by a nucleic acid molecule that hybridizesunder stringent conditions to the opposite strand of a nucleic acidmolecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of anortholog of an amino acid sequence shown in SEQ ID NO:2, wherein saidortholog is encoded by a nucleic acid molecule that hybridizes understringent conditions to the opposite strand of a nucleic acid moleculeshown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequenceshown in SEQ ID NO:2, wherein said fragment comprises at least 10contiguous amino acids.
 2. An isolated peptide comprising an amino acidsequence selected from the group consisting of: (a) an amino acidsequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelicvariant of an amino acid sequence shown in SEQ ID NO:2, wherein saidallelic variant is encoded by a nucleic acid molecule that hybridizesunder stringent conditions to the opposite strand of a nucleic acidmolecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of anortholog of an amino acid sequence shown in SEQ ID NO:2, wherein saidortholog is encoded by a nucleic acid molecule that hybridizes understringent conditions to the opposite strand of a nucleic acid moleculeshown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequenceshown in SEQ ID NO:2, wherein said fragment comprises at least 10contiguous amino acids.
 3. An isolated antibody that selectively bindsto a peptide of claim
 2. 4. An isolated nucleic acid molecule consistingof a nucleotide sequence selected from the group consisting of: (a) anucleotide sequence that encodes an amino acid sequence shown in SEQ IDNO:2; (b) a nucleotide sequence that encodes of an allelic variant of anamino acid sequence shown in SEQ ID NO:2, wherein said nucleotidesequence hybridizes under stringent conditions to the opposite strand ofa nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotidesequence that encodes an ortholog of an amino acid sequence shown in SEQID NO:2, wherein said nucleotide sequence hybridizes under stringentconditions to the opposite strand of a nucleic acid molecule shown inSEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment ofan amino acid sequence shown in SEQ ID NO:2, wherein said fragmentcomprises at least 10 contiguous amino acids; and (e) a nucleotidesequence that is the complement of a nucleotide sequence of (a)-(d). 5.An isolated nucleic acid molecule comprising a nucleotide sequenceselected from the group consisting of: (a) a nucleotide sequence thatencodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotidesequence that encodes of an allelic variant of an amino acid sequenceshown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes understringent conditions to the opposite strand of a nucleic acid moleculeshown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes anortholog of an amino acid sequence shown in SEQ ID NO:2, wherein saidnucleotide sequence hybridizes under stringent conditions to theopposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3;(d) a nucleotide sequence that encodes a fragment of an amino acidsequence shown in SEQ ID NO:2, wherein said fragment comprises at least10 contiguous amino acids; and (e) a nucleotide sequence that is thecomplement of a nucleotide sequence of (a)-(d).
 6. A gene chipcomprising a nucleic acid molecule of claim
 5. 7. A transgenic non-humananimal comprising a nucleic acid molecule of claim
 5. 8. A nucleic acidvector comprising a nucleic acid molecule of claim
 5. 9. A host cellcontaining the vector of claim
 8. 10. A method for producing any of thepeptides of claim 1 comprising introducing a nucleotide sequenceencoding any of the amino acid sequences in (a)-(d) into a host cell,and culturing the host cell under conditions in which the peptides areexpressed from the nucleotide sequence.
 11. A method for producing anyof the peptides of claim 2 comprising introducing a nucleotide sequenceencoding any of the amino acid sequences in (a)-(d) into a host cell,and culturing the host cell under conditions in which the peptides areexpressed from the nucleotide sequence.
 12. A method for detecting thepresence of any of the peptides of claim 2 in a sample, said methodcomprising contacting said sample with a detection agent thatspecifically allows detection of the presence of the peptide in thesample and then detecting the presence of the peptide.
 13. A method fordetecting the presence of a nucleic acid molecule of claim 5 in asample, said method comprising contacting the sample with anoligonucleotide that hybridizes to said nucleic acid molecule understringent conditions and determining whether the oligonucleotide bindsto said nucleic acid molecule in the sample.
 14. A method foridentifying a modulator of a peptide of claim 2, said method comprisingcontacting said peptide with an agent and determining if said agent hasmodulated the function or activity of said peptide.
 15. The method ofclaim 14, wherein said agent is administered to a host cell comprisingan expression vector that expresses said peptide.
 16. A method foridentifying an agent that binds to any of the peptides of claim 2, saidmethod comprising contacting the peptide with an agent and assaying thecontacted mixture to determine whether a complex is formed with theagent bound to the peptide.
 17. A pharmaceutical composition comprisingan agent identified by the method of claim 16 and a pharmaceuticallyacceptable carrier therefor.
 18. A method for treating a disease orcondition mediated by a human kinase protein, said method comprisingadministering to a patient a pharmaceutically effective amount of anagent identified by the method of claim
 16. 19. A method for identifyinga modulator of the expression of a peptide of claim 2, said methodcomprising contacting a cell expressing said peptide with an agent, anddetermining if said agent has modulated the expression of said peptide.20. An isolated human kinase peptide having an amino acid sequence thatshares at least 70% homology with an amino acid sequence shown in SEQ IDNO:2.
 21. A peptide according to claim 20 that shares at least 90percent homology with an amino acid sequence shown in SEQ ID NO:2. 22.An isolated nucleic acid molecule encoding a human kinase peptide, saidnucleic acid molecule sharing at least 80 percent homology with anucleic acid molecule shown in SEQ ID NOS:1 or
 3. 23. A nucleic acidmolecule according to claim 22 that shares at least 90 percent homologywith a nucleic acid molecule shown in SEQ ID NOS: 1 or 3.